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Kre29p is a novel nuclear protein involved in DNA repair and mitotic fidelity in Candida glabrata

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Abstract

Candida glabrata KRE29 is an ortholog of Saccharomyces cerevisiae KRE29. S. cerevisiae Kre29p has been identified by affinity purification as a subunit of the Smc5–Smc6 complex, which is required for DNA repair and chromosome segregation. However, mutant phenotypes of S. cerevisiae KRE29 have not been well characterized and none of its orthologs’ functions has been reported. Here we report phenotypic characteristics of a C. glabrata kre29 deletant. The absence of C. glabrata Kre29p resulted in decreased viability, exhibiting cell cycle arrest between late S-phase and metaphase even under normal growth conditions, and also caused an increase of plasmid loss rate, implying that Kre29p is required for mitotic chromosome transmission fidelity. The deletant showed increased sensitivity to high temperature as well as to DNA damaging agents including UV, gamma ray, 4-nitroquinoline-1-oxide and methyl methanesulfonate, and the phenotypes were restored in the KRE29 reintegrant. Consistent with the Δkre29 phenotypes, a Kre29p-GFP fusion protein was located in the nucleus. Furthermore, Kre29p-GFP became concentrated and formed distinct foci after exposure to 4-nitroquinoline-1-oxide. These results suggest the involvement of C. glabrata Kre29p in DNA repair. To our knowledge, this is the first report addressing a cellular protein involved in DNA repair in C. glabrata.

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References

  • Bodey GP, et al (2002) The epidemiology of Candida glabrata and Candida albicans fungemia in immunocompromised patients with cancer. Am J Med 112:380–385

    Article  PubMed  Google Scholar 

  • Cormack BP, Bertram G, Egerton M, Gow NA, Falkow S, Brown AJ (1997) Yeast-enhanced green fluorescent protein (yEGFP)a reporter of gene expression in Candida albicans. Microbiology 143(Pt 2):303–311

    Article  PubMed  CAS  Google Scholar 

  • Cormack BP, Falkow S (1999) Efficient homologous and illegitimate recombination in the opportunistic yeast pathogen Candida glabrata. Genetics 151:979–987

    PubMed  CAS  Google Scholar 

  • Domergue R, et al (2005) Nicotinic acid limitation regulates silencing of Candida adhesins during UTI. Science 308:866–870

    Article  PubMed  CAS  Google Scholar 

  • Espinel-Ingroff A, et al (1992) Collaborative comparison of broth macrodilution and microdilution antifungal susceptibility tests. J Clin Microbiol 30:3138–3145

    PubMed  CAS  Google Scholar 

  • Ferreira MG, Cooper JP (2004) Two modes of DNA double-strand break repair are reciprocally regulated through the fission yeast cell cycle. Genes Dev 18:2249–2254

    Article  PubMed  CAS  Google Scholar 

  • Fidel PL Jr, Vazquez JA, Sobel JD (1999) Candida glabrata: review of epidemiology, pathogenesis, and clinical disease with comparison to C. albicans. Clin Microbiol Rev 12:80–96

    PubMed  Google Scholar 

  • Fonzi WA, Irwin MY (1993) Isogenic strain construction and gene mapping in Candida albicans. Genetics 134:717–728

    PubMed  CAS  Google Scholar 

  • Frieman MB, McCaffery JM, Cormack BP (2002) Modular domain structure in the Candida glabrata adhesin Epa1p, a beta1,6 glucan-cross-linked cell wall protein. Mol Microbiol 46:479–492

    Article  PubMed  CAS  Google Scholar 

  • Fujioka Y, Kimata Y, Nomaguchi K, Watanabe K, Kohno K (2002) Identification of a novel non-structural maintenance of chromosomes (SMC) component of the SMC5-SMC6 complex involved in DNA repair. J Biol Chem 277:21585–21591

    Article  PubMed  CAS  Google Scholar 

  • Geber A, et al (1995) Deletion of the Candida glabrata ERG3 and ERG11 genes: effect on cell viability, cell growth, sterol composition, and antifungal susceptibility. Antimicrob Agents Chemother 39:2708–2717

    PubMed  CAS  Google Scholar 

  • Giaever G, et al (2002) Functional profiling of the Saccharomyces cerevisiae genome. Nature 418:387–391

    Article  PubMed  CAS  Google Scholar 

  • Gow NA, et al (1994) A hyphal-specific chitin synthase gene (CHS2) is not essential for growth, dimorphism, or virulence of Candida albicans. Proc Natl Acad Sci USA 91:6216–6220

    Article  PubMed  CAS  Google Scholar 

  • Haase SB, Lew DJ (1997) Flow cytometric analysis of DNA content in budding yeast. Methods Enzymol 283:322–332

    Article  PubMed  CAS  Google Scholar 

  • Hazbun TR, et al (2003) Assigning function to yeast proteins by integration of technologies. Mol Cell 12:1353–1365

    Article  PubMed  CAS  Google Scholar 

  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2:163–167

    Article  PubMed  CAS  Google Scholar 

  • Hu B, et al (2005) Qri2/Nse4, a component of the essential Smc5/6 DNA repair complex. Mol Microbiol 55:1735–1750

    Article  PubMed  CAS  Google Scholar 

  • Izumikawa K, Kakeya H, Tsai HF, Grimberg B, Bennett JE (2003) Function of Candida glabrata ABC transporter gene, PDH1. Yeast 20:249–261

    Article  PubMed  CAS  Google Scholar 

  • Jessberger R (2002) The many functions of SMC proteins in chromosome dynamics. Nat Rev Mol Cell Biol 3:767–778

    Article  PubMed  CAS  Google Scholar 

  • Kaye JA, Melo JA, Cheung SK, Vaze MB, Haber JE, Toczyski DP (2004) DNA breaks promote genomic instability by impeding proper chromosome segregation. Curr Biol 14:2096–2106

    Article  PubMed  CAS  Google Scholar 

  • Kitada K, Yamaguchi E, Arisawa M (1996) Isolation of a Candida glabrata centromere and its use in construction of plasmid vectors. Gene 175:105–108

    Article  PubMed  CAS  Google Scholar 

  • Koshland D, Rutledge L, Fitzgerald-Hayes M, Hartwell LH (1987) A genetic analysis of dicentric minichromosomes in Saccharomyces cerevisiae. Cell 48:801–812

    Article  PubMed  CAS  Google Scholar 

  • Lee SE, Moore JK, Holmes A, Umezu K, Kolodner RD, Haber JE (1998) Saccharomyces Ku70, mre11/rad50 and RPA proteins regulate adaptation to G2/M arrest after DNA damage. Cell 94:399–409

    Article  PubMed  CAS  Google Scholar 

  • Lehmann AR (2005) The role of SMC proteins in the responses to DNA damage. DNA Repair (Amst) 4:309–314

    Article  CAS  Google Scholar 

  • Lisby M, Mortensen UH, Rothstein R (2003) Colocalization of multiple DNA double-strand breaks at a single Rad52 repair centre. Nat Cell Biol 5:572–577

    Article  PubMed  CAS  Google Scholar 

  • Lisby M, Rothstein R (2004) DNA repair: keeping it together. Curr Biol 14:R994–R996

    Article  PubMed  CAS  Google Scholar 

  • Lisby M, Rothstein R, Mortensen UH (2001) Rad52 forms DNA repair and recombination centers during S phase. Proc Natl Acad Sci USA 98:8276–8282

    Article  PubMed  CAS  Google Scholar 

  • Losada A, Hirano T (2005) Dynamic molecular linkers of the genome: the first decade of SMC proteins. Genes Dev 19:1269–1287

    Article  PubMed  CAS  Google Scholar 

  • Mehta S, Yang XM, Jayaram M, Velmurugan S (2005) A novel role for the mitotic spindle during DNA segregation in yeast: promoting 2 microm plasmid-cohesin association. Mol Cell Biol 25:4283–4298

    Article  PubMed  CAS  Google Scholar 

  • Onoda F, et al (2004) SMC6 is required for MMS-induced interchromosomal and sister chromatid recombinations in Saccharomyces cerevisiae. DNA Repair (Amst) 3:429–439

    Article  CAS  Google Scholar 

  • Page N, et al (2003) A Saccharomyces cerevisiae genome-wide mutant screen for altered sensitivity to K1 killer toxin. Genetics 163:875–894

    PubMed  CAS  Google Scholar 

  • Pfaller MA, Diekema DJ, Jones RN, Messer SA, Hollis RJ (2002) Trends in antifungal susceptibility of Candida spp. isolated from pediatric and adult patients with bloodstream infections: SENTRY antimicrobial surveillance program, 1997 to 2000. J Clin Microbiol 40:852–856

    Article  PubMed  CAS  Google Scholar 

  • Pfaller MA, et al (1999) Trends in species distribution and susceptibility to fluconazole among blood stream isolates of Candida species in the United States. Diagn Microbiol Infect Dis 33:217–222

    Article  PubMed  CAS  Google Scholar 

  • Pierce AJ, Stark JM, Araujo FD, Moynahan ME, Berwick M, Jasin M (2001) Double-strand breaks and tumorigenesis. Trends Cell Biol 11:S52–S59

    PubMed  CAS  Google Scholar 

  • Poddar A, Roy N, Sinha P (1999) MCM21 and MCM22, two novel genes of the yeast Saccharomyces cerevisiae are required for chromosome transmission. Mol Microbiol 31:349–360

    Article  PubMed  CAS  Google Scholar 

  • Ross KE, Cohen-Fix O (2003) The role of Cdh1p in maintaining genomic stability in budding yeast. Genetics 165:489–503

    PubMed  CAS  Google Scholar 

  • Sambrook J, Russell W (2001) Molecular cloning: a laboratory manual. 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Sandell LL, Zakian VA (1993) Loss of a yeast telomere: arrest, recovery, and chromosome loss. Cell 75:729–739

    Article  PubMed  CAS  Google Scholar 

  • Seno JD, Dynlacht JR (2004) Intracellular redistribution and modification of proteins of the Mre11/Rad50/Nbs1 DNA repair complex following irradiation and heat-shock. J Cell Physiol 199:157–170

    Article  PubMed  CAS  Google Scholar 

  • Spencer F, Gerring SL, Connelly C, Hieter P (1990) Mitotic chromosome transmission fidelity mutants in Saccharomyces cerevisiae. Genetics 124:237–249

    PubMed  CAS  Google Scholar 

  • Stoyan T, Gloeckner G, Diekmann S, Carbon J (2001) Multifunctional centromere binding factor 1 is essential for chromosome segregation in the human pathogenic yeast Candida glabrata. Mol Cell Biol 21:4875–4888

    Article  PubMed  CAS  Google Scholar 

  • Toczyski DP, Galgoczy DJ, Hartwell LH (1997) CDC5 and CKII control adaptation to the yeast DNA damage checkpoint. Cell 90:1097–1106

    Article  PubMed  CAS  Google Scholar 

  • Torres-Rosell J, et al (2005) SMC5 and SMC6 genes are required for the segregation of repetitive chromosome regions. Nat Cell Biol 7:412–419

    Article  PubMed  CAS  Google Scholar 

  • Tsai HF, et al (2004) Candida glabrata erg1 mutant with increased sensitivity to azoles and to low oxygen tension. Antimicrob Agents Chemother 48:2483–2489

    Article  PubMed  CAS  Google Scholar 

  • Vazquez JA, et al (1999) Evolution of vaginal Candida species recovered from human immunodeficiency virus-infected women receiving fluconazole prophylaxis: the emergence of Candida glabrata? Terry Beirn Community programs for clinical research in AIDS (CPCRA). Clin Infect Dis 28:1025–1031

    Article  PubMed  CAS  Google Scholar 

  • Warren CD, Brady DM, Johnston RC, Hanna JS, Hardwick KG, Spencer FA (2002) Distinct chromosome segregation roles for spindle checkpoint proteins. Mol Biol Cell 13:3029–3041

    Article  PubMed  CAS  Google Scholar 

  • Weinert T (1998) DNA damage and checkpoint pathways: molecular anatomy and interactions with repair. Cell 94:555–558

    Article  PubMed  CAS  Google Scholar 

  • Zhao X, Blobel G (2005) A SUMO ligase is part of a nuclear multiprotein complex that affects DNA repair and chromosomal organization. Proc Natl Acad Sci USA 102:4777–4782

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We are grateful to Orna Cohen-Fix for helpful comments during the project and on the manuscript. We thank Owen Schwartz for confocal microscopy, Yo Hoshino for flow cytometry analysis, Brendan P. Cormack for providing the plasmids (pYGFP1 and pGRB2.2), and William A. Fonzi for the plasmid p5921. This research was supported by the Intramural Research Program of the NIH, National Institute of Allergy and Infectious Diseases.

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  1. Clinical Mycology Section, Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Clinical Center Room 11C304, Bethesda, MD, 20892, USA

    Taiga Miyazaki, Huei-Fung Tsai & John E. Bennett

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  1. Taiga Miyazaki
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  2. Huei-Fung Tsai
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  3. John E. Bennett
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Correspondence to John E. Bennett.

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Communicated by B. Cormack

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Miyazaki, T., Tsai, HF. & Bennett, J.E. Kre29p is a novel nuclear protein involved in DNA repair and mitotic fidelity in Candida glabrata . Curr Genet 50, 11–22 (2006). https://doi.org/10.1007/s00294-006-0072-3

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