Antifungal assay

This application is a continuation of Ser. No. 09/541,782 filed Apr. 3, 2000, now U.S. Pat. No. 6,284,480.

The invention relates to methods for the identification of compounds that modulate the activity of bimC and use of such methods for the identification of therapeutic agents and agricultural compounds.

There is a compelling need to prevent and treat opportunistic fungal infections that result in a variety of manifestations, many of which are fatal if untreated. Indeed, the 1980s witnessed an epidemic rise in Candida infections. A steep rise in the incidence of Aspergillus infections has occurred during the 1990's. Similar rises in zygomycosis, cryptococcosis, histoplasmosis and fusaria infection have also been noted. The reasons for the rise in fungal infections are several, but a key factor is the growing population of immuno-compromised individuals. This group includes patients with HIV disease (AIDS), older patients, patients who have undergone invasive surgery, transplant patients and burn victims.

Accordingly, the need for novel antifungal drugs is clear. At present there are two accepted therapies, namely, amphotericin B and the azole family of compounds. Both are effective for certain fungal infections but each have drawbacks-amphotericin B is nephrotoxic and there are increasing numbers of azole resistant strains. These drugs act by either directly or indirectly disrupting the fungal cell wall (or its underlying membrane), which has been the target of choice for antifungal agents.

As the population of immunosuppressed individuals increases, so do the numbers and types of fungal infections noted in these patients. Although candidiasis remains the most common fungal infection in immunosuppressed patients, aspergillosis, zygomycosis, and other filamentous fungal infections are a major problem for an increasing number of patients. The endemic mycoses, especially histoplasmosis and coccidioidomycosis, also constitute a risk for patients. Those at particular risk for such infections are those with AIDS, those having undergone bone marrow or organ transplants, those receiving chemotherapy and others who have had debilitating illness, severe injury, prolonged hospitalization, or long-term treatment with antibacterial drugs.

According to the CDC's National Nosocomial Surveillance System, the rate of hospital-related fungal infections nearly doubled between 1980 and 1990. In 1997, an estimated 240,000 individuals showed clinical symptoms of endemic mycoses. With the current approaches to treatment (primarily amphotericin B and the azoles) the mortality rate in patients with systemic fungal infections ranges from 30-100%, depending on the infection.

The severity of the fungal infections increases as the immune system becomes more dysfunctional. Fungi are among the most ubiquitous pathogens seen in patients with AIDS; virtually all major fungal pathogens cause disease in HIV-positive patients. The majority of untreated HIV-positive patients experience at least one episode of fungal infection and many fungal infections are AIDS-defining illnesses in AIDS patients.

Many emerging fungal pathogens are resistant to the currently available antifungal agents and, thus, pose a special risk for immunocompromised patients. Furthermore, as existing therapies are limited in the range of fungal pathogens against which they show efficacy, and as identification of the fungal pathogen infecting an individual is slow relative to the urgent need to control the infection, there is a critical need to develop broad-spectrum antifungal therapeutics.

Clinically relevant mycotic diseases include candidiasis, aspergillosis, cryptococcosis, endemic mycoses and infections caused by a host of new, emerging fungal pathogens. The standard of care for the more common mycoses employs amphotericin B and the triazole family of therapeutics. These treatments are far from ideal; substantial side effects are seen with amphotericin B. For the azoles problematic issues include increasingly resistant strains and limited bioavailability. Finally, for some of the endemic mycoses as well as new emerging fungal pathogens, no appropriate therapeutics are available. The best prospect for combating existing and emerging threats is through the development of broad-spectrum antifungals.

Given the increased numbers of patients manifesting mycotic diseases, combined with the lack of effective antifungals and the increase in azole resistant isolates, there is a compelling need for novel antifungals. The present invention provides a novel method to identify such antifungals.

The present invention provides methods to identify candidate agents that bind to a target protein or act as a modulator of the binding characteristics or biological activity of a target protein. In one embodiment, the method is performed in plurality simultaneously. For example, the method can be performed at the same time on multiple assay mixtures in a multi-well screening plate. Furthermore, in a preferred embodiment, fluorescence or absorbance readouts are utilized to determine activity. Thus, in one aspect, the invention provides a high throughput screening system for detecting modulators of bimC enzyme activity.

Preferably, the target protein either directly or indirectly produces ADP or phosphate. More preferably, the target protein comprises bimC or a fragment thereof.

The method further comprises adding a candidate agent to a mixture comprising the target protein under conditions that normally allow the production of ADP or phosphate. The method further comprises subjecting the mixture to an enzymatic reaction that uses said ADP or phosphate as a substrate under conditions that normally allow the ADP or phosphate to be utilized and determining the level of activity of the enzymatic reaction as a measure of the concentration of ADP or phosphate. The phrase “use ADP or phosphate” means that the ADP or phosphate are directly acted upon by detection reagents. In one case, the ADP, for example, can be hydrolyzed or can be phosphorylated. As another example, the phosphate can be added to another compound. As used herein, in each of these cases, ADP or phosphate is acting as a substrate. A change in the level between the presence and absence of the candidate agent indicates a modulator of the target protein.

Also provided are modulators of the bimC subfamily of kinesins including antifungal agents. The agents and compositions provided herein can be used in variety of applications which include the formulation of sprays, powders, and other compositions in the formulation of antifungal preparations which can be used to prevent the growth of fungi. Also provided herein are methods of preventing and treating fungal infections.

FIGS. 1A and 1B show the nucleic acid sequence of an embodiment of bimC (SEQ ID NO:1).

FIG. 2 shows the amino acid sequence of an embodiment of bimC (SEQ ID NO:2).

FIGS. 3A and 3B show the nucleic acid sequence of a bimC homolog, cin8 (SEQ ID NO:3).

FIG. 4 shows the amino acid sequence of cin8 (SEQ ID NO:4).

FIGS. 5A and 5B show the nucleic acid sequence of a bimC homolog, cut7 (SEQ ID NO:5).

FIG. 6 shows the amino acid sequence of cut7 (SEQ ID NO:6).

FIGS. 7A and 7B show the nucleic acid sequence of a bimC homolog, KLP61 (SEQ ID NO:7).

FIG. 8 shows the amino acid sequence of KLP61 (SEQ ID NO:8).

FIG. 9 shows the nucleic acid sequence of a bimC homolog, Eg5 (SEQ ID NO:9).

FIG. 10 shows the amino acid sequence of Eg5 (SEQ ID NO:10).

FIG. 11 illustrates the proposed roles for bimC family members during mitosis. The top panel (normal spindle) shows a normal bipolar mitotic spindle at metaphase. The bottom panel (mutant spindle) shows the abnormal morphology of spindles at a comparable mitotic stage in organisms carrying mutations in bimC family motor proteins. BimC family mutants are characterized by the formation of monastral microtubule arrays containing duplicated spindle poles that fail to separate.

FIG. 12 depicts the cloning and expression of A. nidulans bimC motor domain. In Panel A, an overview of the express is shown. The fragment containing the bimC motor domain is indicated by a filled semi-circle. In B, a Coomassie stained gel of the purification of bimC motor domain is shown.

FIG. 13, panel A shows cultures of four ( S. cerevisae, S. pombe, C. albicans, and A. nidulans ) after 2 day incubation in 25 μl volumes at 25° C. In general, fungi are grown by seeding 100-500 ul of each strain into 25 ul of media and incubated at 25° C. for 2-3 days. Compound effects can be monitored microscopically, by Optical density and/or by treatment with the fluorescent viability stain FUN-1 which changes its fluorescence emission based on the metabolic state of the cell. Viability is ascertained by using a 384 pin replicator to transfer approx. 1 μl of cell suspension onto the appropriate solid media and determining the number of colony forming units. Panel B shows the viability of S. cerevisiae cultures after treatment with dilutions of several compounds from a representative biochemical screen.

FIG. 14 shows the results of the testing of representative candidate agents against S. cerevisiae, A. nidulans and mammalian cells. In this experiment fungal cells were incubated with 2-50 μM of each compound and assayed for growth over time at 25° C. The phenotypes shown were obtained after 4 days incubation at 25° C. In these graphs the dotted line represents cell count at various dilutions serving as a measure of cellular viability.

I. Definitions

An “agricultural compound” as used herein refers to a chemical compound that has utility in agriculture and functions to foster food or fiber crop protection or yield improvement. For example, one such compound may serve as a fungicide to control the spreading of plant diseases.

“Cytoskeletal component” denotes any molecule that is found in association with the cellular cytoskeleton, that plays a role in maintaining or regulating the structural integrity of the cytoskeleton, or that mediates or regulates motile/mechanical events mediated by the cytoskeleton. The term includes cytoskeletal polymers (e.g., actin filaments, microtubules, myosin fragments, intermediate filaments), molecular motors, and cytoskeleton associated regulatory proteins (e.g., tropomyosin, alpha-actinin). It is understood that the definitions which apply to the cytoskeletal system apply to embodiments which are drawn to specific components of the cytoskeletal system.

“Cytoskeletal function” refers to biological roles of the cytoskeleton: to provide structural organization (e.g., microvilli, mitotic spindle) and to mediate motile/mechanical events of the cell (e.g., muscle contraction, mitotic contractile ring, pseudopodal movement, active cell surface deformations, vesicle formation and translocation, mitosis (DNA division), cytokinesis (cell division), intracellular (vesicle) transport, cell polarity and organization, cell motility, muscle contractility.

The terms “isolated”, “purified”, or “biologically pure” refer to material which is substantially or essentially free from components which normally accompany it as found in its native state.

“Molecular motor” refers to a cytoskeletal molecule that utilizes chemical energy to produce mechanical force, and drives the motile, mechanical, or structural/organizational properties of the cytoskeleton.

By “ATPase” herein is meant an enzyme that hydrolyzes ATP. ATPases include proteins comprising molecular motors such as kinesins, dyneins, and myosins.

The terms “polypeptide”, “peptide” and “protein” are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residues is an artificial chemical analogue of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers.

“Candidate agent” (used interchangeably herein with “test composition” and “test compound” and “test agent”) refers to a molecule or composition whose effect on the interaction between two or more cytoskeletal components it is desired to assay. The “test composition” can be any molecule or mixture of molecules, optionally in a suitable carrier.

A “therapeutic” as used herein refers to a compound which is believed to be capable of modulating the cytoskeletal system in vivo which can have application in both human and animal disease. Modulation of the cytoskeletal system would be desirable in a number of conditions including, but not limited to: fungal diseases such as aspergillosis, candidiasis, and topical fungal diseases.

II. The Target Protein

A. Overview

The cytoskeleton in mammalian cells is an excellent target for anti-cancer compounds as demonstrated by the effectiveness of taxol and other microtubule poisons. The components of the fungal cytoskeleton, specifically its microtubules and associated proteins, are distinct from their mammalian counterparts. These differences at the primary sequence level and at the structural level may provide the requisite specificity to target the fungal cytoskeleton while leaving the mammalian cytoskeleton unaffected. As an example, the small molecule inhibitor, benomyl, is a potent inhibitor of fungal tubulin, but has no effect on mammalian tubulin. Though benomyl has the specificity required for an effective antifungal, its human toxicity (which is unrelated to its action on microtubules) precludes its use on patients.

B. BimC

Aspergillus nidulans bimC is a kinesin related protein (KRP) whose gene product encodes an essential mitotic KRP. BimC is the founding member of the bimC subfamily of mitotic kinesins (Morris NR, Enos AP. (1992) Trends Genet 8:1 32-7). Mutants in bimC are blocked in mitosis and arrest with duplicated but unseparated spindle poles. (Enos AP, Morris NR. (1990) Cell 60:6 1019-27). Following the discovery that bimC encodes a KRP, several bimC family members have been identified in diverse organisms ranging from yeast to humans. See FIG. 1 . As described below (Table 1), bimC is unique among fungal kinesins in being essential for viability in all fungi in which it has been tested. Although bimC homologs have been found in diverse organisms, the fungal members of this kinesin subgroup form a “fungal-specific cluster”.

In a particularly preferred embodiment, a fragment of the bimC protein comprising its hydrolytically active “motor” domain is used. This motor domain has been cloned and expressed in bacteria such that large quantities of biochemically active, substantially pure protein are available (FIG. 12 ). The expressed protein can be purified by standard chromatographic procedures to yield a purified, biochemically active protein.

C. Cut7

In S. pombe, two loci were identified which, when mutated, block spindle formation. (Hagan I, Yanagida M. (1990) Nature 11(347):6293 563-6) The predicted product of one of these genes, cut7+, contains an amino-terminal domain similar to the kinesin heavy chain motor domain. See FIGS. 5 and 6. The cut7+ gene most closely resembles the Aspergillus nidulans spindle motor gene bimC. Kinesin-related cut7 protein associates with mitotic and meiotic spindles (Hagan and Yanagida, 1992) and mutations in cut7+ affect spindle integrity. (Hagan I, Yanagida M. (1992) Nature 5 356:6364 74-6). In both wild type Aspergillus nidulans and Schizosaccharomyces pombe the intranuclear spindle is bipolar, with microtubules that emanate from spindle pole bodies at either pole interdigitating in a central overlap zone. In bimC and cut7 mutants, microtubule interdigitation does not appear to take place; instead, two unconnected half spindles form and chromosome separation fails.

D. CIN8 and KIP1 ( Saccharomyces cerevisiae )

CIN8 and KIP1 encode polypeptides related to the heavy chain of the microtubule-based force-generating enzyme kinesin, specifically to those within the bimC subfamily (Hoyt, M. A., He, L, Loo, K K, Saunders W S. (1992) J Cell Biol 118:109-120; Roof, D. M., Meluh, P B, Rose MD. (1992) J Cell Biol 118:95-108; and Saunders, W. S. and M. A. Hoyt. (1992) Cell 70:451-458). Cin8p was found to be required for pole separation during mitotic spindle assembly at 37° C. in temperature sensitive mutants, although overproduced Kip1p could suppress the cin8 mutant phenotype. See FIGS. 3 and 4. At lower temperatures, the activity of at least one of these proteins was required for cell viability, indicating that they perform an essential but redundant function. Cin8p was observed to be a component of the mitotic spindle, colocalizing with the microtubules that lie between the poles. Accordingly, Cin8p and Kip1p perform an essential role in the separation of spindle poles during spindle assembly and a major role in spindle elongation.

E. Other Homologs

Outside the fungal realm, bimC homologs have been described in Drosophila melanogaster (KLP61F/KRP130) (see FIGS. 7 and 8 ), Xenopus laevis (Eg5), Homo sapiens (hsEg5/KSP) (see FIGS. 9 and 10) and other organisms (Table 1). In each case, these homologs are required for the formation and maintenance of a normal, bipolar mitotic spindle. It should be noted, however, that these non-fungal homologs are phytogenetically divergent from their fungal counterparts.

F. The Target Protein

In one aspect, the assays or methods provided include the use of bimC. The context in which the bimC is used will indicate whether the sequence is an amino acid sequence, nucleic acid sequence or other. In preferred embodiments, the bimC is isolated from fungi. In one embodiment, bimC as used herein includes bimC homologs. In a preferred embodiment, the bimC homologs are isolated from fungi or yeast. Preferably, the bimC homologs have a sequence or other structural feature that distinguishes the homologs from mammalian homologs.

III. High Throughput Screening

The present invention provides methods to identify candidate agents that bind to a target protein or act as a modulator of the binding characteristics or biological activity of a target protein. In one embodiment, the method is performed in plurality simultaneously. For example, the method can be performed at the same time on multiple assay mixtures in a multi-well screening plate. Furthermore, in a preferred embodiment, fluorescence or absorbance readouts are utilized to determine activity. Thus, in one aspect, the invention provides a high throughput screening system.

More specifically, the present invention includes methods of screening for compounds effective to modulate molecular motor activity. This assay detects modulators of aspects of kinesin motor function ranging from interaction with microtubules to hydrolysis of ATP. Various molecular motor activities are known in the art and include, but are not limited to, the ability to affect ATP hydrolysis, microtubule binding, gliding, and polymerization/depolymerization (effects on microtubule dynamics), binding to other proteins of the spindle, binding to proteins involved in cell-cycle control, or serving as a substrate to other enzymes, such as kinases or proteases, and specific kinesin cellular activities, such as spindle separation.

In a preferred embodiment, this invention provides an effective method for evaluating the activity of bimC or a fragment thereof in a biochemical assay by monitoring its ability to hydrolyze ATP (ATPase activity). Compounds which modulate the activity of bimC can be found by monitoring the production of either ADP or phosphate by a variety of methods.

In a preferred embodiment, activity is measured by the methods disclosed in Ser. No. 09/314,464, filed May 18, 1999, which is incorporated herein by reference in its entirety. These methods are preferably used in multiwell plate formats and are ideally suited for high throughput screening systems to identify lead compounds for medical therapeutic or agricultural uses. More specifically, this assay detects modulators of any aspect of a kinesin motor function ranging from interaction with microtubules to hydrolysis of ATP. The presence of ADP or inorganic phosphate release is used as the readout for protein activity.

The ADP or phosphate level can be monitored using enzyme systems to result in changes in the absorbance or fluorescence of the assay mixture relative to a control mixture to determine if the test compound or mixture of test compounds has an effect on the protein function. This may be done with a single measurement but is preferably done with multiple measurements of the same sample at different times to effect a kinetic profile. In the case of multiple measurements, the absolute rate of the protein activity can be determined, and such measurements have higher specificity particularly in the presence of test compounds that have similar absorbance or fluorescence properties to that of the enzymatic readout. ATP hydrolysis can be measured in a number of spectrophotometric methods.

Methods for ADP determination commonly use the enzyme system pyruvate kinase and lactate dehydrogenase to couple ADP production to the oxidation of NADH. NADH concentration can be monitored by either absorbance ( Proc Natl Acad Sci U S A 1992 89:4884-4887) or fluorescence ( Biochem J 1990 266:611-4). Several variations on this method have been reported (U.S. Pat. No. 4,923,796). See also Nature 1956 178:632; Mol Pharmacol January 1970 6(1):31-40. One method for monitoring phosphate production is to use purine nucleoside phosphorylase to couple the phosphate production to the cleavage of a purine analog which results in either a change in absorbance ( Proc Natl Acad Sci U S A June 1992, 1;89(11):4884-7) or fluorescence ( Biochem J March 1990, 1;266(2):611-4).

With either method, the rate of ATP hydrolysis by the molecular motor of interest can be measured. Test compounds can be assayed in a highly parallel fashion by using multiwell plates by placing the compounds either individually in wells or testing them in mixtures. Assay components including the molecular motors, protein filaments, coupling enzymes and substrates, and ATP can then be added to the wells and the absorbance or fluorescence of each well of the plate can be measured with a plate reader. A compound which modulates the function of the molecular motor is identified by an increase or decrease in the rate of ATP hydrolysis compared to a control assay in the absence of that compound.

For proteins which use ATP, in addition to methods which measure phosphate or ADP, the remaining ATP can be measured using the luciferin-luciferase system ( Anal Biochem. 1971 40:1-17). One specific example of using this method to find inhibitors of a particular ATPase has been patented (U.S. Pat. No. 5,759,795).

The effect of the test compound on the extent of binding between a test compound and bimC is identified as effective if its effect on the extent of binding, rate of ATP hydrolysis, or the like is above a threshold level (e.g., a several-fold difference in binding level between control and experimental samples).

For testing the effects of any inhibitors identified in the above biochemical screens, conditions have been defined that allow the assaying of growth defects on a panel of fungal reference strains in a 384 well format. These experiments can be carried out by seeding small numbers of yeast (100-500 cells/well) in a 25 μl volume/well (FIG. 13 ). Growth is measured microscopically over time as well as by optical density and viability staining. To determine if the inhibitors are fungicidal or fungistatic, dilutions of treated cells can be prepared in 384 well plates and then transferred to solid media using a 384-pin replicator (FIG. 13 ).

IV. Compounds Suitable for Screening

Candidate agents encompass numerous chemical classes, though typically they are organic molecules, preferably small organic molecules having a molecular weight of more than 100 and less than about 2,500 daltons. Candidate agents are obtained from a wide variety of sources including libraries of synthetic or natural compounds. They include peptides, macromolecules, small molecules, chemical and/or biological mixtures, and fungal, bacterial, or algal extracts. Such compounds, or molecules, may be either biological, synthetic organic, or even inorganic compounds, and may be obtained from several sources, including pharmaceutical companies and specialty suppliers of libraries (e.g., combinatorial libraries) of compounds.

Methods of the present invention are well suited for screening libraries of compounds in multiwell plates (e.g., 96-, 384-, or higher density well plates), with a different test compound in each well. In particular, the methods may be employed with combinatorial libraries. A variety of combinatorial libraries of random-sequence oligonucleotides, polypeptides, or synthetic oligomers have been proposed. A number of small-molecule libraries have also been developed.

Combinatorial libraries may be formed by a variety of solution-phase or solid-phase methods in which mixtures of different subunits are added stepwise to growing oligomers or parent compounds, until a desired compound is synthesized. A library of increasing complexity can be formed in this manner, for example, by pooling multiple choices of reagents with each additional subunit step.

The identity of library compounds with desired effects on the target protein can be determined by conventional means, such as iterative synthesis methods in which sublibraries containing known residues in one subunit position only are identified as containing active compounds.

Compounds having the following characteristics have been identified using the methods of the invention: IC 50 values between 15 and 50 μM; low mammalian cellular toxicity (GI 50 >100 μM); and modest fungicidal activity (3-50 μM) against Aspergillus nidulans and Saccharomyces cerevisiae. (See FIG. 14 ).

V. Applications

In a preferred embodiment, the candidate agents modulate bimC. In a particularly preferred embodiment, the candidate agent can be used, for example, to inhibit the growth or spread of fungi, mold, fruit flies, etc. Fungi as used herein includes yeasts, mildews, and rusts. Preferred candidate agents are identified which cause cell death selectively on fungi or insects.

Generally, the compositions and methods provided herein are directed at preventing and treating infections caused by Chytridiomycetes, Hyphochrytridiomycetes, Plasmodiophoromycetes, Oomycetes, Zygomycetes, Ascomycetes, and Basidiomycetes. Fungal infections which can be inhibited or treated with compositions provided herein include but are not limited to: Candidiasis including but not limited to onchomycosis, chronic mucocutaneous candidiasis, oral candidiasis, epiglottistis, esophagitis, gastrointestinal infections, genitourinary infections, for example, caused by any Candida species, including but not limited to Candida albicans, Candida tropicalis, Candida (Torulopsis) glabrata, Candida parapsilosis, Candida lusitaneae, Candida rugosa and Candida pseudotropicalis; Aspergillosis including but not limited to granulocytopenia caused for example, by, Aspergillus spp. including but not limited to A. fumigatus, Aspergillus flavus, Aspergillus niger and Aspergillus terreus; Zygomycosis, including but not limited to pulmonary, sinus and rhinocerebral infections caused by, for example, zygomycetes such as Mucor. Rhizopus spp., Absidia, Rhizomucor, Cunningamella, Saksenaea, Basidobolus and Conidobolus; Cryptococcosis, including but not limited to infections of the central nervous system—meningitis and infections of the respiratory tract caused by, for example, Cryptococcus neoformans; Trichosporonosis caused by, for example, Trichosporon beigelii; Pseudallescheriasis caused by, for example, Pseudallescheria boydii; Fusarium infection caused by, for example, Fusarium such as Fusarium solani, Fusarium moniliforme and Fusarium proliferatum; and other infections such as those caused by, for example, Penicillium spp. (generalized subcutaneous abscesses), Drechslera, Bipolaris, Exserohilum spp., Paecilomyces lilacinum, Exophila jeanselmei (cutaneous nodules), Malassezia furfur (folliculitis), Alternaria (cutaneous nodular lesions), Aureobasidium pullulans (splenic and disseminated infection), Rhodotorula spp. (disseminated infection), Chaetomiuim spp. (empyema), Torulopsis candida (fungernia), Curvularia spp. (nasopharnygeal infection), Cunninghamella spp. (pneumonia), H. Capsulatum, B. dermatitidis, Coccidioides immitis, Sporothrix schenckii and Paracoccidioides brasiliensis, Geotrichum candidum (disseminated infection).

Treating “fungal infections” as used herein refers to the treatment of conditions resulting from fungal infections. Therefore, one may treat, for example, pneumonia, nasopharnygeal infections, disseminated infections and other conditions listed above and known in the art by using the compositions provided herein. In preferred embodiments, treatments and sanitization of areas with the compositions provided herein are provided to immunocompromised patients or areas where there are such patients. Wherein it is desired to identify the particular fungi resulting in the infection, techniques known in the art may be used, see, for example, Musial et al., Clin. Microbiol. Rev., American Society for Microbiology, 349-64 (1998), incorporated herein by reference.

One of skill in the art will readily appreciate that the methods described herein also can be used for diagnostic applications. A diagnostic as used herein is a compound or method that assists in the identification and characterization of a health or disease state in humans or other animals. As described above, bimC is a useful diagnostic tool in vitro for identifying fungal infections.

The present invention also provides for kits for screening for modulators of bimC. Such kits can be prepared from readily available materials and reagents. For example, such kits can comprise any one or more of the following materials: biologically active bimC or a fragment or homolog thereof; reaction tubes; and instructions for testing bimC activity. A wide variety of kits and components can be prepared according to the present invention, depending upon the intended user of the kit and the particular needs of that user.

It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

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1

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A.nidulans

1

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ttctccgcgg cggcagacca aattacggtg tacgaggatg tagttctgcc aattgtcact 600

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aacacccgtg cccagcttga agaggaagag atgctgcgat gtgcgcatga agagactgaa 1800

caccagctcc aggatgtcgg caaaggactt atatcaaccc ttggccaaac cgttgaagat 1860

attaatagcc tacaatcaaa gcttgatcgg aaagccgagt tggacgctac caatgcggaa 1920

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gaggctttcc agacgcggca tgcaaagctt ctcgaaacca cgtctgtcaa agttaacgag 2040

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cgctcgcttg atgcggcatg taacaatgcg aaggctgaga catctagtgc tcacgaagac 2160

atgaacaatg tgcttgaaga aatcaaagat ctgcgcgagg aagtcaagtc taaagtagga 2220

gagggactta atggcctctc agctgccgca gcccggatat cggaggaggt tattggtgaa 2280

ttcacccaac ttcacagcca actgcacaca tccttcaata accttggaaa agacctgaaa 2340

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cgggccgagc tacagagctc gaaccgccag aacatagaaa cgacgcacaa ggcctccgct 2460

catctcgctc aagcgattga agaagaacac gtcgctgcgg aagcggaacg tgagatttta 2520

atgtcacaga tcaaagcgct ggttgaggaa tctcgccaga agcaattcgc ccgcctcagg 2580

gccaagattg acggggtcag gaccgagatt tcagcatcag gggacatgtt agagcaggcc 2640

acaactcagc atgaccgcca gatcgatgag tgggttttca agtctgagca attcgctaag 2700

gatgtcaatg catcgaaaga tgagatcagg acgaagctgc aaaatgattg ggaggcattt 2760

gatcagcgga attcgacaat ccggaaggca acagaatctg tccataagga gacggtacgc 2820

attgttgacg ttcaagtaga cgacatggga cggcaaatgg aagctctgga cgatttcgtg 2880

gcaaaggcac gatctcagaa tggtcgttac cgtgatgcgc atattgcaac cctggataca 2940

atagccaccg gtgttcgcga ctcctactcc tcgattgagg ggcgggttga aaacttgact 3000

ggccggatga accagttcca acaagaagca acccatcatc atgccactct ggaagaatcc 3060

attgctccgc tatcaaacga tgttaggaag ccccttacgg acttgtcttc cagttttcaa 3120

aatcgttctc tggaggagta tgtcgccact ggtgtcaccc cgaagaaacg gaagtacgac 3180

tatatttctg tcttgcctag cacggagtct catgaggtcc tcaagtctcg cctgagaaca 3240

acaaaggaga tggaagtcct tccattcaac agcgacgacc agttgtccgg cccttccagc 3300

tctcccggag gttctccgtc gaaaggcttc gtttacaatg acgtcgagga cgaggtagga 3360

actcatgcac caaccgtgac caacgtcaac ccttccaaca ctggacttag ggaagtcgat 3420

gcaaacgtcg ccgcaagacc gctcgtgtat agcacgggcg agaaatccac ggaccaggat 3480

ggttccccag ttgtcagccc ggatagcgcg acagaagcag aagggatgaa cggaccacca 3540

tccaaaaggc gacgctcaaa ttcagttgtt gctgatacta agttgccaaa caagatgctc 3600

gctaggagga tggctggcat gatggagggg agggaaaacg ttccgcctcc tggtatttcc 3660

aatgggcgcc gactgagggg gcgaccttcg ccctgatata ctcttcactc ggacgagttg 3720

attttctcac cgcgaagtat acttataccc tattgctttt ctcgaggtca tgggtttggc 3780

gttctgggaa caatgagatc tgtatcgggc ttttgataga tcgcatatga tacatcgtct 3840

tcgaaagggt tgggtgcttt tttttttttt tgatcggatt tgttgggcat gtgtttttgg 3900

gtccggtgtg ggcttctctt ctaccaacat gatgtctggc atccttgaga tacctttttt 3960

gtttagtcgg gctacgattt tgacctagat acttttacct atctccgggg aatt 4014

2

1184

A. nidulans

2

Met Ala Gly Pro Gln Arg Ala Thr Tyr Gly Leu Gly Thr Arg Arg Thr

1 5 10 15

Thr Thr Arg Gln Pro Thr Arg Arg Ala Gly Ser Ala Ile Pro Glu Arg

20 25 30

Gln Thr Ser Thr Ala Ser Pro Ala Val Ser Thr Lys Thr Ala Ala Ile

35 40 45

Ser Arg Thr Arg Thr Leu Lys Ser Pro Gly Glu Pro Ala Ser Val Leu

50 55 60

Ala Lys Arg Lys Glu Arg Asp Ile Glu Arg Glu Ile Asn Glu Asp Thr

65 70 75 80

Ser Ile His Val Val Val Arg Cys Arg Gly Arg Asn Glu Arg Glu Val

85 90 95

Lys Glu Asn Ser Gly Val Val Leu Gln Thr Glu Gly Val Lys Gly Lys

100 105 110

Thr Val Glu Leu Ser Met Gly Pro Asn Ala Val Ser Asn Lys Thr Tyr

115 120 125

Thr Phe Asp Lys Val Phe Ser Ala Ala Ala Asp Gln Ile Thr Val Tyr

130 135 140

Glu Asp Val Val Leu Pro Ile Val Thr Glu Met Leu Ala Gly Tyr Asn

145 150 155 160

Cys Thr Ile Phe Ala Tyr Gly Gln Thr Gly Thr Gly Lys Thr Tyr Thr

165 170 175

Met Ser Gly Asp Met Thr Asp Thr Leu Gly Ile Leu Ser Asp Asn Ala

180 185 190

Gly Ile Ile Pro Arg Val Leu Tyr Ser Leu Phe Ala Lys Leu Ala Asp

195 200 205

Thr Glu Ser Thr Val Lys Cys Ser Phe Ile Glu Leu Tyr Asn Glu Glu

210 215 220

Leu Arg Asp Leu Leu Ser Ala Glu Glu Asn Pro Lys Leu Lys Ile Tyr

225 230 235 240

Asp Asn Glu Gln Lys Lys Gly His Met Ser Thr Leu Val Gln Gly Met

245 250 255

Glu Glu Thr Tyr Ile Asp Ser Ala Thr Ala Gly Ile Lys Leu Leu Gln

260 265 270

Gln Gly Ser His Lys Arg Gln Val Ala Ala Thr Lys Cys Asn Asp Leu

275 280 285

Ser Ser Arg Ser His Thr Val Phe Thr Ile Thr Val Asn Ile Lys Arg

290 295 300

Thr Thr Glu Ser Gly Glu Glu Tyr Val Cys Pro Gly Lys Leu Asn Leu

305 310 315 320

Val Asp Leu Ala Gly Ser Glu Asn Ile Gly Arg Ser Gly Ala Glu Asn

325 330 335

Lys Arg Ala Thr Glu Ala Gly Leu Ile Asn Lys Ser Leu Leu Thr Leu

340 345 350

Gly Arg Val Ile Asn Ala Leu Val Asp Lys Ser Gln His Ile Pro Tyr

355 360 365

Arg Glu Ser Lys Leu Thr Arg Leu Leu Gln Asp Ser Leu Gly Gly Arg

370 375 380

Thr Lys Thr Cys Ile Ile Ala Thr Met Ser Pro Ala Arg Ser Asn Leu

385 390 395 400

Glu Glu Thr Ile Ser Thr Leu Asp Tyr Ala Phe Arg Ala Lys Asn Ile

405 410 415

Arg Asn Lys Pro Gln Ile Asn Ser Thr Met Pro Lys Met Thr Leu Leu

420 425 430

Arg Glu Phe Thr Ala Glu Ile Glu Lys Leu Lys Ala Glu Leu Ile Ala

435 440 445

Thr Arg His Arg Asn Gly Val Tyr Met Ser Val Glu Ser Tyr Glu Glu

450 455 460

Met Lys Met Glu Asn Glu Ser Arg Arg Ile Ile Ser Glu Glu Gln Arg

465 470 475 480

Ala Lys Ile Glu Ser Met Glu Ser Ser Leu Arg His Lys Val Gln Glu

485 490 495

Leu Leu Thr Leu Thr Ser Lys Phe Asn Asp Leu Lys Lys Asp Asn Asp

500 505 510

Asp Thr Leu Ala Ala Leu Cys Ser Thr Asn Asp Val Leu Gln Gln Thr

515 520 525

Asp Ile Val Leu Gln Asn Thr Arg Ala Gln Leu Glu Glu Glu Glu Met

530 535 540

Leu Arg Cys Ala His Glu Glu Thr Glu His Gln Leu Gln Asp Val Gly

545 550 555 560

Lys Gly Leu Ile Ser Thr Leu Gly Gln Thr Val Glu Asp Ile Asn Ser

565 570 575

Leu Gln Ser Lys Leu Asp Arg Lys Ala Glu Leu Asp Ala Thr Asn Ala

580 585 590

Glu Leu Trp Arg Ala Ser Ser Thr Glu Val Ser Asp Val Thr Lys Arg

595 600 605

Ile Asp Gln Arg Val Glu Ala Phe Gln Thr Arg His Ala Lys Leu Leu

610 615 620

Glu Thr Thr Ser Val Lys Val Asn Glu Phe Ile Ala Thr Glu Ile Ser

625 630 635 640

Asn Ile Glu Arg Thr Arg Ser Asp Leu Ser Glu Tyr Asn Arg Ser Leu

645 650 655

Asp Ala Ala Cys Asn Asn Ala Lys Ala Glu Thr Ser Ser Ala His Glu

660 665 670

Asp Met Asn Asn Val Leu Glu Glu Ile Lys Asp Leu Arg Glu Glu Val

675 680 685

Lys Ser Lys Val Gly Glu Gly Leu Asn Gly Leu Ser Ala Ala Ala Ala

690 695 700

Arg Ile Ser Glu Glu Val Ile Gly Glu Phe Thr Gln Leu His Ser Gln

705 710 715 720

Leu His Thr Ser Phe Asn Asn Leu Gly Lys Asp Leu Lys Ser Ile Phe

725 730 735

Glu Thr Met Ala Thr His Leu Ser Glu Gln Lys Asn Glu Ile Asn Arg

740 745 750

Leu Arg Ala Glu Leu Gln Ser Ser Asn Arg Gln Asn Ile Glu Thr Thr

755 760 765

His Lys Ala Ser Ala His Leu Ala Gln Ala Ile Glu Glu Glu His Val

770 775 780

Ala Ala Glu Ala Glu Arg Glu Ile Leu Met Ser Gln Ile Lys Ala Leu

785 790 795 800

Val Glu Glu Ser Arg Gln Lys Gln Phe Ala Arg Leu Arg Ala Lys Ile

805 810 815

Asp Gly Val Arg Thr Glu Ile Ser Ala Ser Gly Asp Met Leu Glu Gln

820 825 830

Ala Thr Thr Gln His Asp Arg Gln Ile Asp Glu Trp Val Phe Lys Ser

835 840 845

Glu Gln Phe Ala Lys Asp Val Asn Ala Ser Lys Asp Glu Ile Arg Thr

850 855 860

Lys Leu Gln Asn Asp Trp Glu Ala Phe Asp Gln Arg Asn Ser Thr Ile

865 870 875 880

Arg Lys Ala Thr Glu Ser Val His Lys Glu Thr Val Arg Ile Val Asp

885 890 895

Val Gln Val Asp Asp Met Gly Arg Gln Met Glu Ala Leu Asp Asp Phe

900 905 910

Val Ala Lys Ala Arg Ser Gln Asn Gly Arg Tyr Arg Asp Ala His Ile

915 920 925

Ala Thr Leu Asp Thr Ile Ala Thr Gly Val Arg Asp Ser Tyr Ser Ser

930 935 940

Ile Glu Gly Arg Val Glu Asn Leu Thr Gly Arg Met Asn Gln Phe Gln

945 950 955 960

Gln Glu Ala Thr His His His Ala Thr Leu Glu Glu Ser Ile Ala Pro

965 970 975

Leu Ser Asn Asp Val Arg Lys Pro Leu Thr Asp Leu Ser Ser Ser Phe

980 985 990

Gln Asn Arg Ser Leu Glu Glu Tyr Val Ala Thr Gly Val Thr Pro Lys

995 1000 1005

Lys Arg Lys Tyr Asp Tyr Ile Ser Val Leu Pro Ser Thr Glu Ser His

1010 1015 1020

Glu Val Leu Lys Ser Arg Leu Arg Thr Thr Lys Glu Met Glu Val Leu

1025 1030 1035 1040

Pro Phe Asn Ser Asp Asp Gln Leu Ser Gly Pro Ser Ser Ser Pro Gly

1045 1050 1055

Gly Ser Pro Ser Lys Gly Phe Val Tyr Asn Asp Val Glu Asp Glu Val

1060 1065 1070

Gly Thr His Ala Pro Thr Val Thr Asn Val Asn Pro Ser Asn Thr Gly

1075 1080 1085

Leu Arg Glu Val Asp Ala Asn Val Ala Ala Arg Pro Leu Val Tyr Ser

1090 1095 1100

Thr Gly Glu Lys Ser Thr Asp Gln Asp Gly Ser Pro Val Val Ser Pro

1105 1110 1115 1120

Asp Ser Ala Thr Glu Ala Glu Gly Met Asn Gly Pro Pro Ser Lys Arg

1125 1130 1135

Arg Arg Ser Asn Ser Val Val Ala Asp Thr Lys Leu Pro Asn Lys Met

1140 1145 1150

Leu Ala Arg Arg Met Ala Gly Met Met Glu Gly Arg Glu Asn Val Pro

1155 1160 1165

Pro Pro Gly Ile Ser Asn Gly Arg Arg Leu Arg Gly Arg Pro Ser Pro

1170 1175 1180

3

3884

Saccharomyces cerevisiae

3

aagcaagaat tgaacatgga tgaattcatt ggatcaaaga ccgatttaat caaagatcaa 60

gtgagagata ttcttgataa attgaatatt atttaattct tcatttagaa aaatttcagc 120

tgcttttttt tttctttttc tttccttagg cgtctcgagg ttacaagtcg gagtccctct 180

tcactatcgt ttgtccactt tttttatatc cccattattt tcaatctgaa tttcattttt 240

tttttttaat tcatgaaatt tatatgtccc acgtattact acatatttgc gtttttaatt 300

aaataaataa ctgttacttt tattatatct tatttgcaga tcacttatct gatcaaatgt 360

tttcgttttc gtgtgtggtg acgatgtatt aggtacgcga aataaacaaa acaaacaaac 420

aaggccgcaa caataacatc atctaaagac ttcctttgtg acccgcttct caacagcggg 480

tgtagaactt atggtatggc cagaaagtaa cgttgagtat agatacagaa gcaagcaatt 540

caaaggaaaa agtaataaaa agtatataaa agcgcaaaaa atacaacaag aaagaatttg 600

tttgatgcca gcggaaaacc aaaatacggg tcaagataga agctccaaca gcatcagtaa 660

aaatggcaac tctcaggttg gatgtcacac tgttcctaat gaggaactga acatcactgt 720

agctgtgcga tgcagaggaa ggaatgaaag ggaaattagt atgaaaagct ccgttgtggt 780

aaatgttcca gatattacag gttctaaaga aatttccatt aacacgacgg gagataccgg 840

tataactgct caaatgaatg ccaagagata cacagtggac aaagtcttcg gtcccggcgc 900

ttcccaggat ctaatttttg atgaagtggc gggcccatta ttccaggatt tcattaaagg 960

ttacaattgc accgtactgg tatatggtat gacgtcaaca ggtaaaacat atacaatgac 1020

gggcgacgaa aagttatata atggtgaatt gagcgatgca gcaggaatta taccgagggt 1080

tcttttgaag ttgtttgaca cattggaact acaacagaac gattacgtag taaaatgttc 1140

gttcattgaa ctctacaacg aagaattgaa ggacctcttg gacagcaata gcaacggctc 1200

tagtaatact ggctttgacg gccaatttat gaaaaaattg aggatttttg cttcaagcac 1260

agcaaataat accactagca acagtgctag tagttccagg agtaattcta ggaacagttc 1320

tccgaggtca ttaaatgatc taacacctaa agctgctcta ttaagaaaaa ggttaaggac 1380

aaaatcactg ccgaatacca tcaagcaaca gtatcaacaa caacaggcag tgaattccag 1440

gaacaactct tcctctaact ctggctctac cactaataat gcttctagta acaccaacac 1500

aaataacggt caaagaagtt cgatggctcc aaatgaccaa actaatggta tatacatcca 1560

gaatttgcaa gaatttcaca taacaaatgc tatggagggg ctaaacctat tacaaaaagg 1620

cttaaagcat aggcaagtag cgtccactaa aatgaacgat ttttccagta gatctcatac 1680

catttttaca atcactttgt ataagaagca tcaggatgaa ctatttagaa tttccaaaat 1740

gaatcttgtg gatttagctg gttcagaaaa catcaacaga tccggagcat taaatcaacg 1800

tgccaaagaa gctggttcaa tcaaccaaag tctattgacg ctgggcaggg tcataaacgc 1860

actcgtagat aaaagcggcc atataccttt ccgtgaatcg aaattgaccc gcctgcttca 1920

agattccctg ggtggtaata cgaaaaccgc actaattgct actatatcgc ctgcaaaggt 1980

aacttctgaa gaaacctgca gtacattaga gtatgcttcg aaggctaaaa acattaagaa 2040

caagccgcaa ctgggttcat ttataatgaa ggatattttg gttaaaaata taactatgga 2100

attagcaaag attaaatccg atttactctc tacaaagtcc aaagaaggaa tatatatgag 2160

ccaagatcac tacaaaaatt tgaacagtga tttagaaagt tataaaaatg aagttcaaga 2220

atgtaaaaga gaaattgaaa gtttgacatc gaaaaatgca ttgctagtaa aagataaatt 2280

gaagtcaaaa gaaactattc aatctcaaaa ttgccaaata gaatcattga aaactaccat 2340

agatcattta agggcacaac tagataaaca gcataaaact gaaattgaaa tatccgattt 2400

taataacaaa ctacagaagt tgactgaggt aatgcaaatg gccctacatg attacaaaaa 2460

aagagaactt gaccttaatc aaaagtttga aatgcatatt actaaagaaa ttaaaaaatt 2520

gaaatctaca ctgtttttac aattaaacac tatgcaacag gaaagtattc ttcaagagac 2580

taatatccaa ccaaatcttg atatgatcaa aaatgaagta ctgactctta tgagaaccat 2640

gcaagaaaaa gctgaactaa tgtacaaaga ctgtgtgaag aaaattttaa acgaatctcc 2700

taaattcttc aatgttgtta ttgagaaaat cgacataata agagtagatt tccaaaaatt 2760

ttataaaaat atagccgaga atctttctga tattagcgaa gaaaataaca acatgaaaca 2820

gtacttaaaa aaccattttt tcaagaataa ccatcaagaa ttactgaatc gtcatgtgga 2880

ttctacttat gaaaatattg agaagagaac aaacgagttt gttgagaact ttaaaaaggt 2940

cctaaatgac caccttgacg aaaataaaaa actaataatg cacaatctga caactgcaac 3000

cagcgcggtt attgatcaag aaatggatct gtttgaaccc aagcgcgtta aatgggaaaa 3060

ttcatttgat ctgataaatg attgtgactc catgaataac gaattctata atagcatggc 3120

agcgacgcta tcgcaaatca agagtactgt tgatacatca tcaaattcga tgaatgagtc 3180

tatttcagtc atgaaaggac aagtggaaga atcggagaac gctatatccc ttttgaagaa 3240

caataccaaa tttaatgatc aatttgagca gcttattaac aagcataaca tgttgaaaga 3300

taacattaaa aattcgataa catcaacaca ctctcatata actaatgtgg atgatatcta 3360

taatacgatt gaaaacataa tgaaaaacta tggtaacaag gaaaacgcta ccaaagacga 3420

aatgatcgag aacatattga aggaaatacc aaatctaagt aagaaaatgc cgttaaggtt 3480

atcaaacata aatagcaatt cagtgcaaag tgtaatatcg cccaaaaagc atgcaattga 3540

agatgaaaac aaatccagtg aaaatgtgga caatgagggc tcgagaaaaa tgttaaagat 3600

tgaatagttg atattgcctt tcagtcgaat atatattcaa actagtggtt aataaaaaca 3660

aagtatgtaa agaatactca gttattcatt agaaggcaag acagaagaga agggtgtgaa 3720

accacctcta ccaaacacac caagagatga acctaaatca aattttcaca gagctaacta 3780

tataaacgtt tggattcgtg tgtactatct ttatttacgg aaataagttg taatattaaa 3840

aaaaaaaaaa aacattttga tggacaatga atttctctaa tttt 3884

4

1038

Saccharomyces cerevisiae

4

Met Val Trp Pro Glu Ser Asn Val Glu Tyr Arg Tyr Arg Ser Lys Gln

1 5 10 15

Phe Lys Gly Lys Ser Asn Lys Lys Tyr Ile Lys Ala Gln Lys Ile Gln

20 25 30

Gln Glu Arg Ile Cys Leu Met Pro Ala Glu Asn Gln Asn Thr Gly Gln

35 40 45

Asp Arg Ser Ser Asn Ser Ile Ser Lys Asn Gly Asn Ser Gln Val Gly

50 55 60

Cys His Thr Val Pro Asn Glu Glu Leu Asn Ile Thr Val Ala Val Arg

65 70 75 80

Cys Arg Gly Arg Asn Glu Arg Glu Ile Ser Met Lys Ser Ser Val Val

85 90 95

Val Asn Val Pro Asp Ile Thr Gly Ser Lys Glu Ile Ser Ile Asn Thr

100 105 110

Thr Gly Asp Thr Gly Ile Thr Ala Gln Met Asn Ala Lys Arg Tyr Thr

115 120 125

Val Asp Lys Val Phe Gly Pro Gly Ala Ser Gln Asp Leu Ile Phe Asp

130 135 140

Glu Val Ala Gly Pro Leu Phe Gln Asp Phe Ile Lys Gly Tyr Asn Cys

145 150 155 160

Thr Val Leu Val Tyr Gly Met Thr Ser Thr Gly Lys Thr Tyr Thr Met

165 170 175

Thr Gly Asp Glu Lys Leu Tyr Asn Gly Glu Leu Ser Asp Ala Ala Gly

180 185 190

Ile Ile Pro Arg Val Leu Leu Lys Leu Phe Asp Thr Leu Glu Leu Gln

195 200 205

Gln Asn Asp Tyr Val Val Lys Cys Ser Phe Ile Glu Leu Tyr Asn Glu

210 215 220

Glu Leu Lys Asp Leu Leu Asp Ser Asn Ser Asn Gly Ser Ser Asn Thr

225 230 235 240

Gly Phe Asp Gly Gln Phe Met Lys Lys Leu Arg Ile Phe Ala Ser Ser

245 250 255

Thr Ala Asn Asn Thr Thr Ser Asn Ser Ala Ser Ser Ser Arg Ser Asn

260 265 270

Ser Arg Asn Ser Ser Pro Arg Ser Leu Asn Asp Leu Thr Pro Lys Ala

275 280 285

Ala Leu Leu Arg Lys Arg Leu Arg Thr Lys Ser Leu Pro Asn Thr Ile

290 295 300

Lys Gln Gln Tyr Gln Gln Gln Gln Ala Val Asn Ser Arg Asn Asn Ser

305 310 315 320

Ser Ser Asn Ser Gly Ser Thr Thr Asn Asn Ala Ser Ser Asn Thr Asn

325 330 335

Thr Asn Asn Gly Gln Arg Ser Ser Met Ala Pro Asn Asp Gln Thr Asn

340 345 350

Gly Ile Tyr Ile Gln Asn Leu Gln Glu Phe His Ile Thr Asn Ala Met

355 360 365

Glu Gly Leu Asn Leu Leu Gln Lys Gly Leu Lys His Arg Gln Val Ala

370 375 380

Ser Thr Lys Met Asn Asp Phe Ser Ser Arg Ser His Thr Ile Phe Thr

385 390 395 400

Ile Thr Leu Tyr Lys Lys His Gln Asp Glu Leu Phe Arg Ile Ser Lys

405 410 415

Met Asn Leu Val Asp Leu Ala Gly Ser Glu Asn Ile Asn Arg Ser Gly

420 425 430

Ala Leu Asn Gln Arg Ala Lys Glu Ala Gly Ser Ile Asn Gln Ser Leu

435 440 445

Leu Thr Leu Gly Arg Val Ile Asn Ala Leu Val Asp Lys Ser Gly His

450 455 460

Ile Pro Phe Arg Glu Ser Lys Leu Thr Arg Leu Leu Gln Asp Ser Leu

465 470 475 480

Gly Gly Asn Thr Lys Thr Ala Leu Ile Ala Thr Ile Ser Pro Ala Lys

485 490 495

Val Thr Ser Glu Glu Thr Cys Ser Thr Leu Glu Tyr Ala Ser Lys Ala

500 505 510

Lys Asn Ile Lys Asn Lys Pro Gln Leu Gly Ser Phe Ile Met Lys Asp

515 520 525

Ile Leu Val Lys Asn Ile Thr Met Glu Leu Ala Lys Ile Lys Ser Asp

530 535 540

Leu Leu Ser Thr Lys Ser Lys Glu Gly Ile Tyr Met Ser Gln Asp His

545 550 555 560

Tyr Lys Asn Leu Asn Ser Asp Leu Glu Ser Tyr Lys Asn Glu Val Gln

565 570 575

Glu Cys Lys Arg Glu Ile Glu Ser Leu Thr Ser Lys Asn Ala Leu Leu

580 585 590

Val Lys Asp Lys Leu Lys Ser Lys Glu Thr Ile Gln Ser Gln Asn Cys

595 600 605

Gln Ile Glu Ser Leu Lys Thr Thr Ile Asp His Leu Arg Ala Gln Leu

610 615 620

Asp Lys Gln His Lys Thr Glu Ile Glu Ile Ser Asp Phe Asn Asn Lys

625 630 635 640

Leu Gln Lys Leu Thr Glu Val Met Gln Met Ala Leu His Asp Tyr Lys

645 650 655

Lys Arg Glu Leu Asp Leu Asn Gln Lys Phe Glu Met His Ile Thr Lys

660 665 670

Glu Ile Lys Lys Leu Lys Ser Thr Leu Phe Leu Gln Leu Asn Thr Met

675 680 685

Gln Gln Glu Ser Ile Leu Gln Glu Thr Asn Ile Gln Pro Asn Leu Asp

690 695 700

Met Ile Lys Asn Glu Val Leu Thr Leu Met Arg Thr Met Gln Glu Lys

705 710 715 720

Ala Glu Leu Met Tyr Lys Asp Cys Val Lys Lys Ile Leu Asn Glu Ser

725 730 735

Pro Lys Phe Phe Asn Val Val Ile Glu Lys Ile Asp Ile Ile Arg Val

740 745 750

Asp Phe Gln Lys Phe Tyr Lys Asn Ile Ala Glu Asn Leu Ser Asp Ile

755 760 765

Ser Glu Glu Asn Asn Asn Met Lys Gln Tyr Leu Lys Asn His Phe Phe

770 775 780

Lys Asn Asn His Gln Glu Leu Leu Asn Arg His Val Asp Ser Thr Tyr

785 790 795 800

Glu Asn Ile Glu Lys Arg Thr Asn Glu Phe Val Glu Asn Phe Lys Lys

805 810 815

Val Leu Asn Asp His Leu Asp Glu Asn Lys Lys Leu Ile Met His Asn

820 825 830

Leu Thr Thr Ala Thr Ser Ala Val Ile Asp Gln Glu Met Asp Leu Phe

835 840 845

Glu Pro Lys Arg Val Lys Trp Glu Asn Ser Phe Asp Leu Ile Asn Asp

850 855 860

Cys Asp Ser Met Asn Asn Glu Phe Tyr Asn Ser Met Ala Ala Thr Leu

865 870 875 880

Ser Gln Ile Lys Ser Thr Val Asp Thr Ser Ser Asn Ser Met Asn Glu

885 890 895

Ser Ile Ser Val Met Lys Gly Gln Val Glu Glu Ser Glu Asn Ala Ile

900 905 910

Ser Leu Leu Lys Asn Asn Thr Lys Phe Asn Asp Gln Phe Glu Gln Leu

915 920 925

Ile Asn Lys His Asn Met Leu Lys Asp Asn Ile Lys Asn Ser Ile Thr

930 935 940

Ser Thr His Ser His Ile Thr Asn Val Asp Asp Ile Tyr Asn Thr Ile

945 950 955 960

Glu Asn Ile Met Lys Asn Tyr Gly Asn Lys Glu Asn Ala Thr Lys Asp

965 970 975

Glu Met Ile Glu Asn Ile Leu Lys Glu Ile Pro Asn Leu Ser Lys Lys

980 985 990

Met Pro Leu Arg Leu Ser Asn Ile Asn Ser Asn Ser Val Gln Ser Val

995 1000 1005

Ile Ser Pro Lys Lys His Ala Ile Glu Asp Glu Asn Lys Ser Ser Glu

1010 1015 1020

Asn Val Asp Asn Glu Gly Ser Arg Lys Met Leu Lys Ile Glu

1025 1030 1035

5

4223

S.pombe

5

aacggaagaa atcttgctgc ttagcttctt cgacaactgg agcaccttcg attactacaa 60

ccgtgtcata tccaacaggt ttagttacat taagcttaac ttccagttca gaaacatcga 120

tttcctcgtc ctttacgacg atttgatcct taatttcctc aattaggatt tccgacattt 180

ctggtgtagt gccgttgtct ctagtcaagt ttcttcaccc agtgactggg gggacatcga 240

agagtaactt tagagaaatt taaaaaaaat tgaaagtaat gaaaagtaga atttaaaaaa 300

catggtatca tttaaaataa actaataaag ttgattctgt tttatatttc ttctttttta 360

gattttaact cattctaccg attttgacta ttatcattac tgacaacagt ttatctgcca 420

agttcgttca ttttgtattg atttttagta ttagcacaat tgttggattt ttcataaaag 480

ttcttttttc tcctttttat actgttgatt tgtactgcaa aacattggaa tgagacattc 540

caagtgttac cgtgtctgag tcctcgttct tgtttacaac aattcaacaa catactcagg 600

agcgaacaat tgtccaattt attcattctt tttaccccta aaaacagacg tttacgatct 660

tttcctattt taaatttacc cctttttttt atattgtggt ttattgaact ttacaacaga 720

attgcaattt aaattaacaa ccttgtcgct ttctttgtaa caatccttac tcataaaacc 780

gaatggcccc tagagttgca cctggtggct cacaacaatt tctaggaaag caggggttga 840

aagctaaaaa cccggtttct actccaaatt cacatttccg tctgcgagca atcctaggaa 900

acgacgtgag cctcctacta ttgacactgg gtacccagat cgctccgaca ctaattctcc 960

gacagatcat gcacttcatg atgagaacga aaccaatatt aacgtagtcg tccgtgttcg 1020

tggtcgtaca gaccaagaag tacgcgacaa tagtagcctt gctgtatcga cttcgggcgc 1080

tatgggtgct gaattggcca tccagtcgga tccttcatcc atgttggtca cgaagacgta 1140

tgcctttgat aaagtctttg gtcctgaagc tgaccagtta atgctattcg aaaattcggt 1200

agcgcccatg ctggagcaag tattgaacgg ctataattgt actatatttg cgtacggaca 1260

aactggaacc ggtaaaacat atactatgtc tggtgatctt agcgattccg atggaatttt 1320

gtctgaagga gcaggcctta ttcctcgtgc cctttatcaa ttattttcct ccttggacaa 1380

cagtaatcaa gaatatgctg tcaaatgctc ctattatgaa ctttacaacg aggaaattcg 1440

cgatcttttg gtttccgaag aattgcgaaa gccagctcgt gtttttgaag atacttcgcg 1500

gagagggaac gttgttatta ctggtattga agaaagttat ataaaaaatg ctggcgatgg 1560

gttgcgattg ttacgtgaag gctctcaccg gcgtcaagtt gctgctacaa agtgtaacga 1620

tctttcatct cgtagtcatt caatcttcac gataactctg catagaaagg tatcttcggg 1680

aatgacagat gaaactaatt cacttaccat aaataataac tctgatgact tgctgcgcgc 1740

cagcaaactt catatggttg atttggctgg aagcgagaac attggacgtt ctggagctga 1800

aaacaaacgt gctcgagaaa ctggaatgat caatcaatca ctattaacat taggccgagt 1860

aattaatgca ttagtggaaa aggcccacca tattccatac cgtgagtcca agcttacacg 1920

cctattgcag gattctttgg ggggaaagac taaaacctct atgatcgtta ccgtatcttc 1980

tactaatact aatttggaag aaacaatttc tactttggag tatgctgctc gtgctaaaag 2040

tatacgcaat aagcctcaaa ataatcagct tgtctttcga aaagtcctta taaaagactt 2100

agtgttagat atcgaacgat tgaaaaatga ccttaatgcc acccgtaaaa agaacggagt 2160

ttatcttgct gaaagcacgt ataaggagct tatggatcgt gttcaaaaca aagatttgtt 2220

atgccaggaa caggctcgca aacttgaagt gctggactta aacgttaaaa gttcgaggga 2280

acaactgcag tatgtttcta aatctaatca agaacataaa aaggaagttg aagctctgca 2340

actgcaactg gttaattcat ctacagagtt agaaagtgta aagtctgaga acgaaaagct 2400

gaaaaatgaa ctagttttag aaatcgaaaa acgaaaaaaa tacgaaacca atgaagctaa 2460

aattacaact gttgcaacag acctttccca atattaccga gaatcaaaag aatacattgc 2520

tagcttatat gagaagcttg atcgcactga acgaaataat aaagaaaatg aaaacaattt 2580

ttggaatctt aagttcaatt tgttaacgat gttgagatct tttcatggaa gttttactga 2640

tgaaacaaat ggctatttta ctttgttgaa tgattttaat gcttcaatgg aagaattatt 2700

gaacacccac agtaatcaac tgctgatttc tatgaccaaa attactgagc attttcagag 2760

tttagatgag gcgttgcaaa gtgctcgttc ttcttgtgca gtacctaatt ctagcttaga 2820

cctaattgtt tctgaattaa aggattccaa aaacagcctt cttgatgcgt tggagcacag 2880

tctgcaggat attagcatgt cttcccagaa gttgggaaat ggaatttctt cggaattgat 2940

agaattgcag aaagacatga aagaatcata ccgacaactt gttcaggaat taagatcact 3000

atataattta cagcatactc atgaagaatc acagaaggag ctaatgtatg gtgtgcgtaa 3060

cgatattgat gctctggtta aaacttgcac aacatctttg aacgatgcag atataatatt 3120

aagtgattac atatctgatc aaaaatccaa atttgaatcc aagcaacaag atttgattgc 3180

taatattggt aaaattgttt caaatttttt acaggaacag aacgaatctt tgtataccaa 3240

agcggatatc ttacattcac atctcaatga tacaaactcg aatataagga aagcaaacga 3300

aattatgaat aaccgttcag aagagttttt acggaatgcg gcttcacaag cagaaatcgt 3360

gggtgccaat aaagaaagga ttcaaaagac agttgaaaat ggatctcaac tgcttgacag 3420

taaaagcaag gccattcata gcaattccag atcaatgtat gaccattgct tggctttagc 3480

ggagtctcaa aaacaaggtg ttaatcttga agttcaaacg ctggatcgtt tgttgcagaa 3540

agtaaaagag cattcagaag ataataccaa ggagaagcat caacaattac ttgatttatt 3600

ggaatccctt gttggcaaca atgacaatct tatcgattcc atcaagacac cgcacactga 3660

attacagaaa attacagatc atgttttaaa gggaacgaca tcacttgcta atcatactaa 3720

tgaattactt ggtttaggag atgaatctct atgtaacctt gaaactacta tagaagacac 3780

gtctttggtg aagctggaaa caactggcga tacaccttcc aaacgagaac ttcctgccac 3840

tccatcttgg acacgagatt cgtctttaat taaggaaact acgaatttaa atttagattc 3900

ggataagaaa ttcgttaggg aaacctacac atcgtctaat caaactaatg agccggacgt 3960

atatgataaa ccatctaatt catcaagaac tagtcttttg cggagtagca gaagtgccta 4020

ttccaaaatg aaacgataat gacacgattt tattaattta tacttaatgg acgaaaatac 4080

aatagaaatg tattacatct tggggctttg gctggttaaa acggaaagtt tggttatgtg 4140

tgtacatact gcgttttaaa taatagtttt aggagttgta tacatataaa gattaattta 4200

gatagaagtt taatgatata cag 4223

6

1073

S.pombe

6

Met Ala Pro Arg Val Ala Pro Gly Gly Ser Gln Gln Phe Leu Gly Lys

1 5 10 15

Gln Gly Leu Lys Ala Lys Asn Pro Val Ser Thr Pro Asn Ser His Phe

20 25 30

Arg Leu Arg Ala Ile Leu Gly Asn Asp Val Ser Leu Leu Leu Leu Thr

35 40 45

Leu Asp His Ala Leu His Asp Glu Asn Glu Thr Asn Ile Asn Val Val

50 55 60

Val Arg Val Arg Gly Arg Thr Asp Gln Glu Val Arg Asp Asn Ser Ser

65 70 75 80

Leu Ala Val Ser Thr Ser Gly Ala Met Gly Ala Glu Leu Ala Ile Gln

85 90 95

Ser Asp Pro Ser Ser Met Leu Val Thr Lys Thr Tyr Ala Phe Asp Lys

100 105 110

Val Phe Gly Pro Glu Ala Asp Gln Leu Met Leu Phe Glu Asn Ser Val

115 120 125

Ala Pro Met Leu Glu Gln Val Leu Asn Gly Tyr Asn Cys Thr Ile Phe

130 135 140

Ala Tyr Gly Gln Thr Gly Thr Gly Lys Thr Tyr Thr Met Ser Gly Asp

145 150 155 160

Leu Ser Asp Ser Asp Gly Ile Leu Ser Glu Gly Ala Gly Leu Ile Pro

165 170 175

Arg Ala Leu Tyr Gln Leu Phe Ser Ser Leu Asp Asn Ser Asn Gln Glu

180 185 190

Tyr Ala Val Lys Cys Ser Tyr Tyr Glu Leu Tyr Asn Glu Glu Ile Arg

195 200 205

Asp Leu Leu Val Ser Glu Glu Leu Arg Lys Pro Ala Arg Val Phe Glu

210 215 220

Asp Thr Ser Arg Arg Gly Asn Val Val Ile Thr Gly Ile Glu Glu Ser

225 230 235 240

Tyr Ile Lys Asn Ala Gly Asp Gly Leu Arg Leu Leu Arg Glu Gly Ser

245 250 255

His Arg Arg Gln Val Ala Ala Thr Lys Cys Asn Asp Leu Ser Ser Arg

260 265 270

Ser His Ser Ile Phe Thr Ile Thr Leu His Arg Lys Val Ser Ser Gly

275 280 285

Met Thr Asp Glu Thr Asn Ser Leu Thr Ile Asn Asn Asn Ser Asp Asp

290 295 300

Leu Leu Arg Ala Ser Lys Leu His Met Val Asp Leu Ala Gly Ser Glu

305 310 315 320

Asn Ile Gly Arg Ser Gly Ala Glu Asn Lys Arg Ala Arg Glu Thr Gly

325 330 335

Met Ile Asn Gln Ser Leu Leu Thr Leu Gly Arg Val Ile Asn Ala Leu

340 345 350

Val Glu Lys Ala His His Ile Pro Tyr Arg Glu Ser Lys Leu Thr Arg

355 360 365

Leu Leu Gln Asp Ser Leu Gly Gly Lys Thr Lys Thr Ser Met Ile Val

370 375 380

Thr Val Ser Ser Thr Asn Thr Asn Leu Glu Glu Thr Ile Ser Thr Leu

385 390 395 400

Glu Tyr Ala Ala Arg Ala Lys Ser Ile Arg Asn Lys Pro Gln Asn Asn

405 410 415

Gln Leu Val Phe Arg Lys Val Leu Ile Lys Asp Leu Val Leu Asp Ile

420 425 430

Glu Arg Leu Lys Asn Asp Leu Asn Ala Thr Arg Lys Lys Asn Gly Val

435 440 445

Tyr Leu Ala Glu Ser Thr Tyr Lys Glu Leu Met Asp Arg Val Gln Asn

450 455 460

Lys Asp Leu Leu Cys Gln Glu Gln Ala Arg Lys Leu Glu Val Leu Asp

465 470 475 480

Leu Asn Val Lys Ser Ser Arg Glu Gln Leu Gln Tyr Val Ser Lys Ser

485 490 495

Asn Gln Glu His Lys Lys Glu Val Glu Ala Leu Gln Leu Gln Leu Val

500 505 510

Asn Ser Ser Thr Glu Leu Glu Ser Val Lys Ser Glu Asn Glu Lys Leu

515 520 525

Lys Asn Glu Leu Val Leu Glu Ile Glu Lys Arg Lys Lys Tyr Glu Thr

530 535 540

Asn Glu Ala Lys Ile Thr Thr Val Ala Thr Asp Leu Ser Gln Tyr Tyr

545 550 555 560

Arg Glu Ser Lys Glu Tyr Ile Ala Ser Leu Tyr Glu Lys Leu Asp Arg

565 570 575

Thr Glu Arg Asn Asn Lys Glu Asn Glu Asn Asn Phe Trp Asn Leu Lys

580 585 590

Phe Asn Leu Leu Thr Met Leu Arg Ser Phe His Gly Ser Phe Thr Asp

595 600 605

Glu Thr Asn Gly Tyr Phe Thr Leu Leu Asn Asp Phe Asn Ala Ser Met

610 615 620

Glu Glu Leu Leu Asn Thr His Ser Asn Gln Leu Leu Ile Ser Met Thr

625 630 635 640

Lys Ile Thr Glu His Phe Gln Ser Leu Asp Glu Ala Leu Gln Ser Ala

645 650 655

Arg Ser Ser Cys Ala Val Pro Asn Ser Ser Leu Asp Leu Ile Val Ser

660 665 670

Glu Leu Lys Asp Ser Lys Asn Ser Leu Leu Asp Ala Leu Glu His Ser

675 680 685

Leu Gln Asp Ile Ser Met Ser Ser Gln Lys Leu Gly Asn Gly Ile Ser

690 695 700

Ser Glu Leu Ile Glu Leu Gln Lys Asp Met Lys Glu Ser Tyr Arg Gln

705 710 715 720

Leu Val Gln Glu Leu Arg Ser Leu Tyr Asn Leu Gln His Thr His Glu

725 730 735

Glu Ser Gln Lys Glu Leu Met Tyr Gly Val Arg Asn Asp Ile Asp Ala

740 745 750

Leu Val Lys Thr Cys Thr Thr Ser Leu Asn Asp Ala Asp Ile Ile Leu

755 760 765

Ser Asp Tyr Ile Ser Asp Gln Lys Ser Lys Phe Glu Ser Lys Gln Gln

770 775 780

Asp Leu Ile Ala Asn Ile Gly Lys Ile Val Ser Asn Phe Leu Gln Glu

785 790 795 800

Gln Asn Glu Ser Leu Tyr Thr Lys Ala Asp Ile Leu His Ser His Leu

805 810 815

Asn Asp Thr Asn Ser Asn Ile Arg Lys Ala Asn Glu Ile Met Asn Asn

820 825 830

Arg Ser Glu Glu Phe Leu Arg Asn Ala Ala Ser Gln Ala Glu Ile Val

835 840 845

Gly Ala Asn Lys Glu Arg Ile Gln Lys Thr Val Glu Asn Gly Ser Gln

850 855 860

Leu Leu Asp Ser Lys Ser Lys Ala Ile His Ser Asn Ser Arg Ser Met

865 870 875 880

Tyr Asp His Cys Leu Ala Leu Ala Glu Ser Gln Lys Gln Gly Val Asn

885 890 895

Leu Glu Val Gln Thr Leu Asp Arg Leu Leu Gln Lys Val Lys Glu His

900 905 910

Ser Glu Asp Asn Thr Lys Glu Lys His Gln Gln Leu Leu Asp Leu Leu

915 920 925

Glu Ser Leu Val Gly Asn Asn Asp Asn Leu Ile Asp Ser Ile Lys Thr

930 935 940

Pro His Thr Glu Leu Gln Lys Ile Thr Asp His Val Leu Lys Gly Thr

945 950 955 960

Thr Ser Leu Ala Asn His Thr Asn Glu Leu Leu Gly Leu Gly Asp Glu

965 970 975

Ser Leu Cys Asn Leu Glu Thr Thr Ile Glu Asp Thr Ser Leu Val Lys

980 985 990

Leu Glu Thr Thr Gly Asp Thr Pro Ser Lys Arg Glu Leu Pro Ala Thr

995 1000 1005

Pro Ser Trp Thr Arg Asp Ser Ser Leu Ile Lys Glu Thr Thr Asn Leu

1010 1015 1020

Asn Leu Asp Ser Asp Lys Lys Phe Val Arg Glu Thr Tyr Thr Ser Ser

1025 1030 1035 1040

Asn Gln Thr Asn Glu Pro Asp Val Tyr Asp Lys Pro Ser Asn Ser Ser

1045 1050 1055

Arg Thr Ser Leu Leu Arg Ser Ser Arg Ser Ala Tyr Ser Lys Met Lys

1060 1065 1070

Arg

7

3709

Drosophila melanogaster

7

tttttttttc agggtgccgt gttgcggtca cactatttgc gcattatttt aaaattgaac 60

ggacaactga agacgctgct tctctctgtt ctcattggaa atagtgcgaa aattggtttt 120

ccacgccagc cttctgtaaa tagtagttac agttatcagc attcggctcg gtggaaggtg 180

aagcacaagg atttccagga atcaaacaca cacacacacg cataccgata cacgcgacag 240

gcgtcatttt ttggactgta cgcgctaaaa ccacaattaa ttgtttcaat tgatcaatat 300

ggacatatct ggtgggaata cgtcgcgcca gccccaaaag aagtccaacc aaaacatcca 360

ggtgtatgtg cgcgtcagac cccttaattc tcgggaacgt tgcatccgct cggccgaagt 420

cgtggatgtg gtcggaccac gggaagtggt cacccgccac acgctggact ccaagctcac 480

caagaagttc acctttgacc gcagttttgg ccccgagtcc aagcagtgcg atgtctactc 540

cgtcgtggtg tctccgctga tcgaggaggt cctcaatggc tataactgca cggtgtttgc 600

ttatggccag acgggcacag ggaagaccca caccatggtg ggcaacgaga ctgccgaact 660

gaaatcctcc tgggaagatg actctgacat tggcatcata ccgcgcgctc tgagtcacct 720

tttcgatgag ctgcgcatga tggaggtgga gtacactatg cgcatttcct acttggaact 780

gtacaatgag gagctgtgcg atctactgtc caccgatgac accaccaaga tacgcatttt 840

cgatgacagc accaagaagg gatcggtgat tatccagggc ctggaggaga taccagtgca 900

cagcaaggat gatgtgtaca agctgctgga gaagggaaag gagcgtcgca aaacagccac 960

tacgctgatg aatgcacagt cctcacgctc ccacactgta ttttctatag ttgtgcacat 1020

cagggagaat ggcatcgaag gagaggacat gctgaaaatc ggtaaactga atctggtgga 1080

tctggcgggc agtgaaaatg tttccaaggc tgggaatgaa aagggaatac gtgtgcggga 1140

aacagtaaac atcaatcaga gcctcttgac tctcggtaga gtaattaccg ctttggtcga 1200

tcgcgctcct cacgttccat atcgtgaatc aaagctgact agactgctgc aagaatctct 1260

gggtggcagg accaagacct ccatcatagc caccatatcg ccgggccaca aggacatcga 1320

ggagaccctg agcactttag agtacgctca tcgcgccaag aacattcaaa acaagcccga 1380

agtcaatcag aagctgacca aaaagactgt gctcaaggaa tacaccgaag aaatcgacaa 1440

gcttaaaaga gatcttatgg cggccaggga caagaatggc atctatttgg ccgaggaaac 1500

ctacggggag ataactttga agttagaatc ccagaaccgc gagcttaacg agaaaatgct 1560

gctgctcaag gctttgaagg acgaactcca gaacaaggag aagatcttca gtgaggtgag 1620

catgagtctt gtggagaaaa cgcaggagct gaaaaagacc gaggagaacc tactgaacac 1680

gaagggtact ttgctcctga ccaagaaagt gctgaccaag accaagcgac gatacaagga 1740

aaaaaaggag ttggtggcct cgcatatgaa aacggagcag gttttgacca cacaggctca 1800

ggagatccta gccgctgctg atctggccac tgatgatact catcagctgc acggaaccat 1860

cgaaagaagg cgtgagttgg atgagaagat tcgccgatct tgcgatcagt tcaaagatcg 1920

tatgcaggat aatttggaaa tgatcggtgg cagcctgaac ttgtatcagg atcagcaggc 1980

agccctcaag gagcagctaa gccaggaaat ggtaaactcg agctatgtga gccaacgcct 2040

tgcactaaat tccagcaaaa gcatcgaaat gcttaaggag atgtgtgccc aatcgctaca 2100

ggatcagacc aacctccata ataagttaat aggagaagtc atgaagatca gcgatcagca 2160

ctctcaggca tttgtcgcca agctgatgga gcagatgcag cagcaacagc ttttgatgag 2220

caaagaaatt cagactaacc tgcaggtgat cgaggaaaac aaccagcggc acaaagccat 2280

gctggactct atgcaggaaa agttcgccac cataattgac agcagcttac agtctgtgga 2340

agaacatgcg aagcaaatgc acaagaagtt ggagcagctc ggggcaatga gtttgccaga 2400

tgcggaggaa ctgcagaatc ttcaagagga gttggcgaat gaacgagctc tggcgcagca 2460

ggaggatgct ctccttgagt ccatgatgat gcagatggaa cagatcaaga acttgcggtc 2520

caaaaatagc atcagcatgt ccgtacatct taataagatg gaggagagtc gactgacgag 2580

aaatcatcgc atcgatgata ttaagtctgg tatccaagac taccaaaagt tgggcataga 2640

agcttcccaa tccgcacaag cggagctcac cagccaaatg gaggcgggaa tgctttgtct 2700

ggaccaaggc gttgccaact gttcgatgct tcaggtgcac atgaagaatc tcaaccagaa 2760

atacgaaaag gaaacaaatg agaatgttgg ttccgttcga gtgcaccaca atcaggtgga 2820

aattatttgc caagagagca agcagcagct tgaggcggtg caggagaaaa ccgaagttaa 2880

cttggaacag atggtggatg caaggcagca gcttatcacc gaggacagac agcgattcat 2940

aggccatgcc actgtagcca ctgatcttgt ccaagagtcg aaccggcagt tctcggagca 3000

tgctgaacac caacggcagc agctgcaaat ttgcgagcaa gagcttgtac gcttccaaca 3060

gtcggagttg aaaacgtacg ccccaactgg gaccacgccc tccaaaaggg atttcgttta 3120

tccgcgcacc ctcgtggcca cgtcgccaca tcaggaaatt gtgaggcgct atcgccaaga 3180

gcaggactgg tcagatctgg acaccacggc cactatagat gagtgcagcg agggtgagca 3240

tgacgactcc atgcattccg tccaggagct gtccgaaact gaaaccataa tgaactccac 3300

gcccattgag cccgtggatg gtgtcaccgt gaagcgtggg tgcggcacca cccgcaattc 3360

caattcgaac gccctaaagc cacccgtagc tactggtgga aaacgcagca gctcgttgtc 3420

acgttccctg acacccagta aaacgtcacc tcgaggttct cccgccttcg tcaggcacaa 3480

caaagaaaac gtagcctgat tgcattcgat gacagtagta gtctcccctt cccagtaacc 3540

cgtttatgtt agtggatttc ggatcgctgt gtttgtttgg ctcttctgga tattcatttt 3600

atctgtatat caatctttgg aatttcgtac atttattatt taaataaact cagtatgcta 3660

tgtaaagtta aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaa 3709

8

1066

Drosophila melanogaster

8

Met Asp Ile Ser Gly Gly Asn Thr Ser Arg Gln Pro Gln Lys Lys Ser

1 5 10 15

Asn Gln Asn Ile Gln Val Tyr Val Arg Val Arg Pro Leu Asn Ser Arg

20 25 30

Glu Arg Cys Ile Arg Ser Ala Glu Val Val Asp Val Val Gly Pro Arg

35 40 45

Glu Val Val Thr Arg His Thr Leu Asp Ser Lys Leu Thr Lys Lys Phe

50 55 60

Thr Phe Asp Arg Ser Phe Gly Pro Glu Ser Lys Gln Cys Asp Val Tyr

65 70 75 80

Ser Val Val Val Ser Pro Leu Ile Glu Glu Val Leu Asn Gly Tyr Asn

85 90 95

Cys Thr Val Phe Ala Tyr Gly Gln Thr Gly Thr Gly Lys Thr His Thr

100 105 110

Met Val Gly Asn Glu Thr Ala Glu Leu Lys Ser Ser Trp Glu Asp Asp

115 120 125

Ser Asp Ile Gly Ile Ile Pro Arg Ala Leu Ser His Leu Phe Asp Glu

130 135 140

Leu Arg Met Met Glu Val Glu Tyr Thr Met Arg Ile Ser Tyr Leu Glu

145 150 155 160

Leu Tyr Asn Glu Glu Leu Cys Asp Leu Leu Ser Thr Asp Asp Thr Thr

165 170 175

Lys Ile Arg Ile Phe Asp Asp Ser Thr Lys Lys Gly Ser Val Ile Ile

180 185 190

Gln Gly Leu Glu Glu Ile Pro Val His Ser Lys Asp Asp Val Tyr Lys

195 200 205

Leu Leu Glu Lys Gly Lys Glu Arg Arg Lys Thr Ala Thr Thr Leu Met

210 215 220

Asn Ala Gln Ser Ser Arg Ser His Thr Val Phe Ser Ile Val Val His

225 230 235 240

Ile Arg Glu Asn Gly Ile Glu Gly Glu Asp Met Leu Lys Ile Gly Lys

245 250 255

Leu Asn Leu Val Asp Leu Ala Gly Ser Glu Asn Val Ser Lys Ala Gly

260 265 270

Asn Glu Lys Gly Ile Arg Val Arg Glu Thr Val Asn Ile Asn Gln Ser

275 280 285

Leu Leu Thr Leu Gly Arg Val Ile Thr Ala Leu Val Asp Arg Ala Pro

290 295 300

His Val Pro Tyr Arg Glu Ser Lys Leu Thr Arg Leu Leu Gln Glu Ser

305 310 315 320

Leu Gly Gly Arg Thr Lys Thr Ser Ile Ile Ala Thr Ile Ser Pro Gly

325 330 335

His Lys Asp Ile Glu Glu Thr Leu Ser Thr Leu Glu Tyr Ala His Arg

340 345 350

Ala Lys Asn Ile Gln Asn Lys Pro Glu Val Asn Gln Lys Leu Thr Lys

355 360 365

Lys Thr Val Leu Lys Glu Tyr Thr Glu Glu Ile Asp Lys Leu Lys Arg

370 375 380

Asp Leu Met Ala Ala Arg Asp Lys Asn Gly Ile Tyr Leu Ala Glu Glu

385 390 395 400

Thr Tyr Gly Glu Ile Thr Leu Lys Leu Glu Ser Gln Asn Arg Glu Leu

405 410 415

Asn Glu Lys Met Leu Leu Leu Lys Ala Leu Lys Asp Glu Leu Gln Asn

420 425 430

Lys Glu Lys Ile Phe Ser Glu Val Ser Met Ser Leu Val Glu Lys Thr

435 440 445

Gln Glu Leu Lys Lys Thr Glu Glu Asn Leu Leu Asn Thr Lys Gly Thr

450 455 460

Leu Leu Leu Thr Lys Lys Val Leu Thr Lys Thr Lys Arg Arg Tyr Lys

465 470 475 480

Glu Lys Lys Glu Leu Val Ala Ser His Met Lys Thr Glu Gln Val Leu

485 490 495

Thr Thr Gln Ala Gln Glu Ile Leu Ala Ala Ala Asp Leu Ala Thr Asp

500 505 510

Asp Thr His Gln Leu His Gly Thr Ile Glu Arg Arg Arg Glu Leu Asp

515 520 525

Glu Lys Ile Arg Arg Ser Cys Asp Gln Phe Lys Asp Arg Met Gln Asp

530 535 540

Asn Leu Glu Met Ile Gly Gly Ser Leu Asn Leu Tyr Gln Asp Gln Gln

545 550 555 560

Ala Ala Leu Lys Glu Gln Leu Ser Gln Glu Met Val Asn Ser Ser Tyr

565 570 575

Val Ser Gln Arg Leu Ala Leu Asn Ser Ser Lys Ser Ile Glu Met Leu

580 585 590

Lys Glu Met Cys Ala Gln Ser Leu Gln Asp Gln Thr Asn Leu His Asn

595 600 605

Lys Leu Ile Gly Glu Val Met Lys Ile Ser Asp Gln His Ser Gln Ala

610 615 620

Phe Val Ala Lys Leu Met Glu Gln Met Gln Gln Gln Gln Leu Leu Met

625 630 635 640

Ser Lys Glu Ile Gln Thr Asn Leu Gln Val Ile Glu Glu Asn Asn Gln

645 650 655

Arg His Lys Ala Met Leu Asp Ser Met Gln Glu Lys Phe Ala Thr Ile

660 665 670

Ile Asp Ser Ser Leu Gln Ser Val Glu Glu His Ala Lys Gln Met His

675 680 685

Lys Lys Leu Glu Gln Leu Gly Ala Met Ser Leu Pro Asp Ala Glu Glu

690 695 700

Leu Gln Asn Leu Gln Glu Glu Leu Ala Asn Glu Arg Ala Leu Ala Gln

705 710 715 720

Gln Glu Asp Ala Leu Leu Glu Ser Met Met Met Gln Met Glu Gln Ile

725 730 735

Lys Asn Leu Arg Ser Lys Asn Ser Ile Ser Met Ser Val His Leu Asn

740 745 750

Lys Met Glu Glu Ser Arg Leu Thr Arg Asn His Arg Ile Asp Asp Ile

755 760 765

Lys Ser Gly Ile Gln Asp Tyr Gln Lys Leu Gly Ile Glu Ala Ser Gln

770 775 780

Ser Ala Gln Ala Glu Leu Thr Ser Gln Met Glu Ala Gly Met Leu Cys

785 790 795 800

Leu Asp Gln Gly Val Ala Asn Cys Ser Met Leu Gln Val His Met Lys

805 810 815

Asn Leu Asn Gln Lys Tyr Glu Lys Glu Thr Asn Glu Asn Val Gly Ser

820 825 830

Val Arg Val His His Asn Gln Val Glu Ile Ile Cys Gln Glu Ser Lys

835 840 845

Gln Gln Leu Glu Ala Val Gln Glu Lys Thr Glu Val Asn Leu Glu Gln

850 855 860

Met Val Asp Ala Arg Gln Gln Leu Ile Thr Glu Asp Arg Gln Arg Phe

865 870 875 880

Ile Gly His Ala Thr Val Ala Thr Asp Leu Val Gln Glu Ser Asn Arg

885 890 895

Gln Phe Ser Glu His Ala Glu His Gln Arg Gln Gln Leu Gln Ile Cys

900 905 910

Glu Gln Glu Leu Val Arg Phe Gln Gln Ser Glu Leu Lys Thr Tyr Ala

915 920 925

Pro Thr Gly Thr Thr Pro Ser Lys Arg Asp Phe Val Tyr Pro Arg Thr

930 935 940

Leu Val Ala Thr Ser Pro His Gln Glu Ile Val Arg Arg Tyr Arg Gln

945 950 955 960

Glu Gln Asp Trp Ser Asp Leu Asp Thr Thr Ala Thr Ile Asp Glu Cys

965 970 975

Ser Glu Gly Glu His Asp Asp Ser Met His Ser Val Gln Glu Leu Ser

980 985 990

Glu Thr Glu Thr Ile Met Asn Ser Thr Pro Ile Glu Pro Val Asp Gly

995 1000 1005

Val Thr Val Lys Arg Gly Cys Gly Thr Thr Arg Asn Ser Asn Ser Asn

1010 1015 1020

Ala Leu Lys Pro Pro Val Ala Thr Gly Gly Lys Arg Ser Ser Ser Leu

1025 1030 1035 1040

Ser Arg Ser Leu Thr Pro Ser Lys Thr Ser Pro Arg Gly Ser Pro Ala

1045 1050 1055

Phe Val Arg His Asn Lys Glu Asn Val Ala

1060 1065

9

3741

H.sapiens

9

gaattccgtc atggcgtcgc agccaaattc gtctgcgaag aagaaagagg agaaggggaa 60

gaacatccag gtggtggtga gatgcagacc atttaatttg gcagagcgga aagctagcgc 120

ccattcaata gtagaatgtg atcctgtacg aaaagaagtt agtgtacgaa ctggaggatt 180

ggctgacaag agctcaagga aaacatacac ttttgatatg gtgtttggag catctactaa 240

acagattgat gtttaccgaa gtgttgtttg tccaattctg gatgaagtta ttatgggcta 300

taattgcact atctttgcgt atggccaaac tggcactgga aaaactttta caatggaagg 360

tgaaaggtca cctaatgaag agtatacctg ggaagaggat cccttggctg gtataattcc 420

acgtaccctt catcaaattt ttgagaaact tactgataat ggtactgaat tttcagtcaa 480

agtgtctctg ttggagatct ataatgaaga gctttttgat cttcttaatc catcatctga 540

tgtttctgag agactacaga tgtttgatga tccccgtaac aagagaggag tgataattaa 600

aggtttagaa gaaattacag tacacaacaa ggatgaagtc tatcaaattt tagaaaaggg 660

ggcagcaaaa aggacaactg cagctactct gatgaatgca tactctagtc gttcccactc 720

agttttctct gttacaatac atatgaaaga aactacgatt gatggagaag agcttgttaa 780

aatcggaaag ttgaacttgg ttgatcttgc aggaagtgaa aacattggcc gttctggagc 840

tgttgataag agagctcggg aagctggaaa tataaatcaa tccctgttga ctttgggaag 900

ggtcattact gcccttgtag aaagaacacc tcatgttcct tatcgagaat ctaaactaac 960

tagaatcctc caggattctc ttggagggcg tacaagaaca tctataattg caacaatttc 1020

tcctgcatct ctcaatcttg aggaaactct gagtacattg gaatatgctc atagagcaaa 1080

gaacatattg aataagcctg aagtgaatca gaaactcacc aaaaaagctc ttattaagga 1140

gtatacggag gagatagaac gtttaaaacg agatcttgct gcagcccgtg agaaaaatgg 1200

agtgtatatt tctgaagaaa attttagagt catgagtgga aaattaactg ttcaagaaga 1260

gcagattgta gaattgattg aaaaaattgg tgctgttgag gaggagctga atagggttac 1320

agagttgttt atggataata aaaatgaact tgaccagtgt aaatctgacc tgcaaaataa 1380

aacacaagaa cttgaaacca ctcaaaaaca tttgcaagaa actaaattac aacttgttaa 1440

agaagaatat atcacatcag ctttggaaag tactgaggag aaacttcatg atgctgccag 1500

caagctgctt aacacagttg aagaaactac aaaagatgta tctggtctcc attccaaact 1560

ggatcgtaag aaggcagttg accaacacaa tgcagaagct caggatattt ttggcaaaaa 1620

cctgaatagt ctgtttaata atatggaaga attaattaag gatggcagct caaagcaaaa 1680

ggccatgcta gaagtacata agaccttatt tggtaatctg ctgtcttcca gtgtctctgc 1740

attagatacc attactacag tagcacttgg atctctcaca tctattccag aaaatgtgtc 1800

tactcatgtt tctcagattt ttaatatgat actaaaagaa caatcattag cagcagaaag 1860

taaaactgta ctacaggaat tgattaatgt actcaagact gatcttctaa gttcactgga 1920

aatgatttta tccccaactg tggtgtctat actgaaaatc aatagtcaac taaagcatat 1980

tttcaagact tcattgacag tggccgataa gatagaagat caaaaaaaaa ggaactcaga 2040

tggctttctc agtatactgt gtaacaatct acatgaacta caagaaaata ccatttgttc 2100

cttggttgag tcacaaaagc aatgtggaaa cctaactgaa gacctgaaga caataaagca 2160

gacccattcc caggaacttt gcaagttaat gaatctttgg acagagagat tctgtgcttt 2220

ggaggaaaag tgtgaaaata tacagaaacc acttagtagt gtccaggaaa atatacagca 2280

gaaatctaag gatatagtca acaaaatgac ttttcacagt caaaaatttt gtgctgattc 2340

tgatggcttc tcacaggaac tcagaaattt taaccaagaa ggtacaaaat tggttgaaga 2400

atctgtgaaa cactctgata aactcaatgg caacctggaa aaaatatctc aagagactga 2460

acagagatgt gaatctctga acacaagaac agtttatttt tctgaacagt gggtatcttc 2520

cttaaatgaa agggaacagg aacttcacaa cttattggag gttgtaagcc aatgttgtga 2580

ggcttcaagt tcagacatca ctgagaaatc agatggacgt aaggcagctc atgagaaaca 2640

gcataacatt tttcttgatc agatgactat tgatgaagat aaattgatag cacaaaatct 2700

agaacttaat gaaaccataa aaattggttt gactaagctt aattgctttc tggaacagga 2760

tctgaaactg gatatcccaa caggtacgac accacagagg aaaagttatt tatacccatc 2820

aacactggta agaactgaac cacgtgaaca tctccttgat cagctgaaaa ggaaacagcc 2880

tgagctgtta atgatgctaa actgttcaga aaacaacaaa gaagagacaa ttccggatgt 2940

ggatgtagaa gaggcagttc tggggcagta tactgaagaa cctctaagtc aagagccatc 3000

tgtagatgct ggtgtggatt gttcatcaat tggcggggtt ccatttttcc agcataaaaa 3060

atcacatgga aaagacaaag aaaacagagg cattaacaca ctggagaggt ctaaagtgga 3120

agaaactaca gagcacttgg ttacaaagag cagattacct ctgcgagccc agatcaacct 3180

ttaattcact tgggggttgg caattttatt tttaaagaaa aacttaaaaa taaaacctga 3240

aaccccagaa cttgagcctt gtgtatagat tttaaaagaa tatatatatc agccgggcgc 3300

gtggctctag ctgtaatccc agctaacttt ggaggctgag gcgggtggat tgcttgagcc 3360

caggagtttg agaccagcct ggccaacgtg cgctaaaacc ttcgtctctg ttaaaaatta 3420

gccgggcgtg gtgggcacac tcctgtaatc ccagctactg gggaggctga ggcacgagaa 3480

tcacttgaac ccagaagcgg ggttgcagtg agccaaaggt acaccactac actccagcct 3540

gggcaacaga gcaagactcg gtctcaaaaa taaaatttaa aaaagatata aggcagtact 3600

gtaaattcag ttgaattttg atatctaccc atttttctgt catccctata gttcactttg 3660

tattaaattg ggtttcattt gggatttgca atgtaaatac gtatttctag ttttcatata 3720

aagtagttct tttaggaatt c 3741

10

1057

H.sapiens

10

Met Ala Ser Gln Pro Asn Ser Ser Ala Lys Lys Lys Glu Glu Lys Gly

1 5 10 15

Lys Asn Ile Gln Val Val Val Arg Cys Arg Pro Phe Asn Leu Ala Glu

20 25 30

Arg Lys Ala Ser Ala His Ser Ile Val Glu Cys Asp Pro Val Arg Lys

35 40 45

Glu Val Ser Val Arg Thr Gly Gly Leu Ala Asp Lys Ser Ser Arg Lys

50 55 60

Thr Tyr Thr Phe Asp Met Val Phe Gly Ala Ser Thr Lys Gln Ile Asp

65 70 75 80

Val Tyr Arg Ser Val Val Cys Pro Ile Leu Asp Glu Val Ile Met Gly

85 90 95

Tyr Asn Cys Thr Ile Phe Ala Tyr Gly Gln Thr Gly Thr Gly Lys Thr

100 105 110

Phe Thr Met Glu Gly Glu Arg Ser Pro Asn Glu Glu Tyr Thr Trp Glu

115 120 125

Glu Asp Pro Leu Ala Gly Ile Ile Pro Arg Thr Leu His Gln Ile Phe

130 135 140

Glu Lys Leu Thr Asp Asn Gly Thr Glu Phe Ser Val Lys Val Ser Leu

145 150 155 160

Leu Glu Ile Tyr Asn Glu Glu Leu Phe Asp Leu Leu Asn Pro Ser Ser

165 170 175

Asp Val Ser Glu Arg Leu Gln Met Phe Asp Asp Pro Arg Asn Lys Arg

180 185 190

Gly Val Ile Ile Lys Gly Leu Glu Glu Ile Thr Val His Asn Lys Asp

195 200 205

Glu Val Tyr Gln Ile Leu Glu Lys Gly Ala Ala Lys Arg Thr Thr Ala

210 215 220

Ala Thr Leu Met Asn Ala Tyr Ser Ser Arg Ser His Ser Val Phe Ser

225 230 235 240

Val Thr Ile His Met Lys Glu Thr Thr Ile Asp Gly Glu Glu Leu Val

245 250 255

Lys Ile Gly Lys Leu Asn Leu Val Asp Leu Ala Gly Ser Glu Asn Ile

260 265 270

Gly Arg Ser Gly Ala Val Asp Lys Arg Ala Arg Glu Ala Gly Asn Ile

275 280 285

Asn Gln Ser Leu Leu Thr Leu Gly Arg Val Ile Thr Ala Leu Val Glu

290 295 300

Arg Thr Pro His Val Pro Tyr Arg Glu Ser Lys Leu Thr Arg Ile Leu

305 310 315 320

Gln Asp Ser Leu Gly Gly Arg Thr Arg Thr Ser Ile Ile Ala Thr Ile

325 330 335

Ser Pro Ala Ser Leu Asn Leu Glu Glu Thr Leu Ser Thr Leu Glu Tyr

340 345 350

Ala His Arg Ala Lys Asn Ile Leu Asn Lys Pro Glu Val Asn Gln Lys

355 360 365

Leu Thr Lys Lys Ala Leu Ile Lys Glu Tyr Thr Glu Glu Ile Glu Arg

370 375 380

Leu Lys Arg Asp Leu Ala Ala Ala Arg Glu Lys Asn Gly Val Tyr Ile

385 390 395 400

Ser Glu Glu Asn Phe Arg Val Met Ser Gly Lys Leu Thr Val Gln Glu

405 410 415

Glu Gln Ile Val Glu Leu Ile Glu Lys Ile Gly Ala Val Glu Glu Glu

420 425 430

Leu Asn Arg Val Thr Glu Leu Phe Met Asp Asn Lys Asn Glu Leu Asp

435 440 445

Gln Cys Lys Ser Asp Leu Gln Asn Lys Thr Gln Glu Leu Glu Thr Thr

450 455 460

Gln Lys His Leu Gln Glu Thr Lys Leu Gln Leu Val Lys Glu Glu Tyr

465 470 475 480

Ile Thr Ser Ala Leu Glu Ser Thr Glu Glu Lys Leu His Asp Ala Ala

485 490 495

Ser Lys Leu Leu Asn Thr Val Glu Glu Thr Thr Lys Asp Val Ser Gly

500 505 510

Leu His Ser Lys Leu Asp Arg Lys Lys Ala Val Asp Gln His Asn Ala

515 520 525

Glu Ala Gln Asp Ile Phe Gly Lys Asn Leu Asn Ser Leu Phe Asn Asn

530 535 540

Met Glu Glu Leu Ile Lys Asp Gly Ser Ser Lys Gln Lys Ala Met Leu

545 550 555 560

Glu Val His Lys Thr Leu Phe Gly Asn Leu Leu Ser Ser Ser Val Ser

565 570 575

Ala Leu Asp Thr Ile Thr Thr Val Ala Leu Gly Ser Leu Thr Ser Ile

580 585 590

Pro Glu Asn Val Ser Thr His Val Ser Gln Ile Phe Asn Met Ile Leu

595 600 605

Lys Glu Gln Ser Leu Ala Ala Glu Ser Lys Thr Val Leu Gln Glu Leu

610 615 620

Ile Asn Val Leu Lys Thr Asp Leu Leu Ser Ser Leu Glu Met Ile Leu

625 630 635 640

Ser Pro Thr Val Val Ser Ile Leu Lys Ile Asn Ser Gln Leu Lys His

645 650 655

Ile Phe Lys Thr Ser Leu Thr Val Ala Asp Lys Ile Glu Asp Gln Lys

660 665 670

Lys Arg Asn Ser Asp Gly Phe Leu Ser Ile Leu Cys Asn Asn Leu His

675 680 685

Glu Leu Gln Glu Asn Thr Ile Cys Ser Leu Val Glu Ser Gln Lys Gln

690 695 700

Cys Gly Asn Leu Thr Glu Asp Leu Lys Thr Ile Lys Gln Thr His Ser

705 710 715 720

Gln Glu Leu Cys Lys Leu Met Asn Leu Trp Thr Glu Arg Phe Cys Ala

725 730 735

Leu Glu Glu Lys Cys Glu Asn Ile Gln Lys Pro Leu Ser Ser Val Gln

740 745 750

Glu Asn Ile Gln Gln Lys Ser Lys Asp Ile Val Asn Lys Met Thr Phe

755 760 765

His Ser Gln Lys Phe Cys Ala Asp Ser Asp Gly Phe Ser Gln Glu Leu

770 775 780

Arg Asn Phe Asn Gln Glu Gly Thr Lys Leu Val Glu Glu Ser Val Lys

785 790 795 800

His Ser Asp Lys Leu Asn Gly Asn Leu Glu Lys Ile Ser Gln Glu Thr

805 810 815

Glu Gln Arg Cys Glu Ser Leu Asn Thr Arg Thr Val Tyr Phe Ser Glu

820 825 830

Gln Trp Val Ser Ser Leu Asn Glu Arg Glu Gln Glu Leu His Asn Leu

835 840 845

Leu Glu Val Val Ser Gln Cys Cys Glu Ala Ser Ser Ser Asp Ile Thr

850 855 860

Glu Lys Ser Asp Gly Arg Lys Ala Ala His Glu Lys Gln His Asn Ile

865 870 875 880

Phe Leu Asp Gln Met Thr Ile Asp Glu Asp Lys Leu Ile Ala Gln Asn

885 890 895

Leu Glu Leu Asn Glu Thr Ile Lys Ile Gly Leu Thr Lys Leu Asn Cys

900 905 910

Phe Leu Glu Gln Asp Leu Lys Leu Asp Ile Pro Thr Gly Thr Thr Pro

915 920 925

Gln Arg Lys Ser Tyr Leu Tyr Pro Ser Thr Leu Val Arg Thr Glu Pro

930 935 940

Arg Glu His Leu Leu Asp Gln Leu Lys Arg Lys Gln Pro Glu Leu Leu

945 950 955 960

Met Met Leu Asn Cys Ser Glu Asn Asn Lys Glu Glu Thr Ile Pro Asp

965 970 975

Val Asp Val Glu Glu Ala Val Leu Gly Gln Tyr Thr Glu Glu Pro Leu

980 985 990

Ser Gln Glu Pro Ser Val Asp Ala Gly Val Asp Cys Ser Ser Ile Gly

995 1000 1005

Gly Val Pro Phe Phe Gln His Lys Lys Ser His Gly Lys Asp Lys Glu

1010 1015 1020

Asn Arg Gly Ile Asn Thr Leu Glu Arg Ser Lys Val Glu Glu Thr Thr

1025 1030 1035 1040

Glu His Leu Val Thr Lys Ser Arg Leu Pro Leu Arg Ala Gln Ile Asn

1045 1050 1055

Leu