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Molecular cloning and analysis of CDC28 and cyclin homologues from the human fungal pathogen Candida albicans

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Abstract

In the budding yeast Saccharomyces cerevisiae, progress of the cell cycle beyond the major control point in G1 phase, termed START, requires activation of the evolutionarily conserved Cdc28 protein kinase by direct association with GI cyclins. We have used a conditional lethal mutation in CDC28 of S. cerevisiae to clone a functional homologue from the human fungal pathogen Candida albicans. The protein sequence, deduced from the nucleotide sequence, is 79% identical to that of S. cerevisiae Cdc28 and as such is the most closely related protein yet identified. We have also isolated from C. albicans two genes encoding putative G1 cyclins, by their ability to rescue a conditional GI cyclin defect in S. cerevisiae; one of these genes encodes a protein of 697 amino acids and is identical to the product of the previously described CCN1 gene. The second gene codes for a protein of 465 residues, which has significant homology to S. cerevisiae Cln3. These data suggest that the events and regulatory mechanisms operating at START are highly conserved between these two organisms.

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References

  • Beach D, Durkacz B, Nurse P (1982) Functionally homologous cell cycle genes in budding and fission yeast. Nature 300:706–709

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acid Res 7:1513–1523

    Google Scholar 

  • Chen M-W, Anne J, Volckaert G, Huysmans E, Vandenberghe A, Wachter RD (1984) The nucleotide sequence of the 5S rRNA of seven molds and a yeast and their use in studying Ascomycete phylogeny. Nucleic Acids Res 12:4881–4892

    Google Scholar 

  • Cryer DR, Eccleshall R, Marmur J (1975) Isolation of yeast DNA. Methods Cell Biol 12:39–44

    Google Scholar 

  • Cvrčková F, Nasmyth K (1993) Yeast G1 cyclins CLN1 and CLN2 and a GAP-like protein have a role in bud formation. EMBO J 13:5277–5286

    Google Scholar 

  • Dower, WJ, Miller, JF, Ragsdale, CW (1988) High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res 16:6127–6145

    Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13

    Google Scholar 

  • Forsburg SL, Nurse P (1991) Identification of a G1-type cyclin puc1+ in the fission yeast Schizosaccharomyces pombe. Nature 351:245–248

    Google Scholar 

  • Forsburg SL, Nurse P (1994) Analysis of the Schizosaccharomyces pombe cyclin pucl: evidence for a role in cell cycle exit. J Cell Sci 107:601–613

    Google Scholar 

  • Hanks SK, Quinn AM (1991) Protein kinase catalytic domain sequence database: identification of conserved features of primary structure and classification of family members. In: Hunter T, Sefton BM (eds) Methods Enzymol 200:38–62

  • Hendriks L, Goris A, Neffs J-M, Peer YVD, Hennebert G, Wachter RD (1989) The nucleotide sequence of the small ribosomal subunit RNA of the yeast Candida albicans and the evolutionary position of the fungi among eukaryotes. Syst Appl Micribiol 13:223–229

    Google Scholar 

  • Humphreys GO, Willshaw GA, Anderson ES (1975) A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta 383:457–463

    Google Scholar 

  • Hurley JL, Donelson JE (1980) Nucleotide sequence of the yeast plasmid. Nature 286:860–865

    Google Scholar 

  • Kurjan J (1993) The pheromone response pathway in Saccharomyces cerevisiae. Annu Rev Genet 27:147–179

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Nasmyth K (1993) Control of the yeast cell cycle by the Cdc28 protein kinase. Curr Biol 5:166–179

    Google Scholar 

  • Nasmyth K, Dirick L (1991) The role of SWI4 and SWI6 in the activity of G1 cyclins in yeast. Cell 66:995–1013

    Google Scholar 

  • Nurse P (1990) Universal control mechanism regulating onset of M-phase. Nature 344:503–508

    Google Scholar 

  • Odds FC (1987) Candida infection: an overview. Crit Rev Microbiol 15:1–5

    Google Scholar 

  • Patterson MN, Sclafani RA, Fangman WL, Rosamond J (1986) Molecular characterization of cell cycle gene CDC7 from Saccharomyces cerevisiae. Mol Cell Biol 6:1590–1598

    Google Scholar 

  • Piggot JR, Rai R, Carter BLA (1982) A bifunctional gene product involved in two phases of the yeast cell cycle. Nature 298:391–393

    Google Scholar 

  • Pringle JR, Hartwell LH (1981) The Saccharomyces cerevisiae cell cycle. In: Strathern JN, Jones EW, Broach JR (eds) The life cycle of the yeast Saccharomyces cerevisiae: life cycle and inheritance, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York pp 97–142

    Google Scholar 

  • Richardson HE, Wittenburg C, Cross F, Reed SI (1989) An essential G1 function for cyclin-like proteins in yeast. Cell 59:1127–1133

    Google Scholar 

  • Rosamond J, Telander KM, Linn S (1979) Modulation of the action of the recBC enzyme of Escherichia coli K-12 by Ca2+. J Biol Chem 254:8646–8652

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Scherer S, Magee PT (1990) Genetics of Candida albicans. Microbiol Rev 54:226–241

    Google Scholar 

  • Schiestl RH, Gietz RD (1989) High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet 16:339–346

    Google Scholar 

  • Sherlock G, Rosamond J (1993) Starting to cycle: G1 controls regulating cell division in budding yeast. J Gen Microbiol 139:2531–2541

    Google Scholar 

  • Sherman F, Fink GR, Hicks (1986) Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Soll DR (1988) High frequency switching in Candida albicans and its relations to vaginal candidiasis. Am J Obstet Gynecol 154:997–1001

    Google Scholar 

  • Tyers M, Tokiwa G, Futcher B (1993) Comparison of the Saccharomyces cerevisiae Gt cyclins: Cln3 may be an upstream activator of Cln1, Cln2 and other cyclins. EMBO J 12:1955–1968

    Google Scholar 

  • Warren G, Sherratt D (1978) Incompatibility and transformation efficiency of ColEl and related plasmids. Mol Gen Genet 161:39–47

    Google Scholar 

  • Whiteway M, Dignard D, Thomas DY (1992) Dominant negative selection of heterologous genes: isolation of Candida albicans genes that interfere with Saccharomyces cerevisiae mating factor-induced cell cycle arrest. Proc Natl Acad Sci USA 89:9410–9414

    Google Scholar 

  • Xiong Y, Connolly T, Futcher B, Beach D (1991) Human D-type cyclin. Cell 65:691–699

    Google Scholar 

  • Zaret K, Sherman F (1982) DNA sequence required for efficient transcription termination in yeast. Cell 28:563–573

    Google Scholar 

  • Zhang H, Scholl R, Browse J, Somerville C (1988) Double stranded DNA sequencing as a choice for DNA sequencing. Nucleic Acids Res 16:1220

    Google Scholar 

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Communicated by B. J. Kilbey

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Sherlock, G., Bahman, A.M., Mahal, A. et al. Molecular cloning and analysis of CDC28 and cyclin homologues from the human fungal pathogen Candida albicans . Molec. Gen. Genet. 245, 716–723 (1994). https://doi.org/10.1007/BF00297278

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