Abstract
Candida albicans is a human fungal pathogen and has been extensively studied because of its clinical importance. Comprehensive gene analyses have, however, made little progress. This is because of the diploid and asexual characteristics of the fungus that hamper gene disruptions. In this study, we found that ultraviolet (UV) irradiation, as well as mutagen treatment, strongly stimulated loss of heterozygosity (LOH) in strains harboring artificially constructed heterozygosity. UV-induced LOH occurred more frequently in cells within the logarithmic phase of growth compared to those within the stationary phase of growth. This was observed at all loci tested on chromosome 7, except for a locus neighboring the centromere. C. albicans RAD52, whose orthologue in Saccharomyces cerevisiae was reported to be involved in DNA repair by homologous recombination, was shown to be required for UV-induced LOH. These results suggest that high efficiency LOH caused by UV irradiation could be a prominent tool for gene analyses in C. albicans.
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References
Berman J, Sudbery PE (2002) Candida albicans: a molecular revolution built on lessons from budding yeast. Nat Rev Genet 3(12):918–930
Chibana H, Magee BB, Grindle S, Ran Y, Scherer S, Magee PT (1998) A physical map of chromosome 7 of Candida albicans. Genetics 149(4):1739–1752
Chibana H, Oka N, Nakayama H, Aoyama T, Magee BB, Magee PT, Mikami Y (2005) Sequence finishing and gene mapping for Candida albicans chromosome 7 and syntenic analysis against the Saccharomyces cerevisiae genome. Genetics 170(4):1525–1537
Ciudad T, Andaluz E, Steinberg-Neifach O, Lue NF, Gow NA, Calderone RA, Larriba G (2004) Homologous recombination in Candida albicans: role of CaRad52p in DNA repair, integration of linear DNA fragments and telomere length. Mol Microbiol 53(4):1177–1194
Daigaku Y, Endo K, Watanabe E, Ono T, Yamamoto K (2004) Loss of heterozygosity and DNA damage repair in Saccharomyces cerevisiae. Mutat Res 556(1–2):183–191
Daigaku Y, Mashiko S, Mishiba K, Yamamura S, Ui A, Enomoto T, Yamamoto K (2006) Loss of heterozygosity in yeast can occur by ultraviolet irradiation during the S phase of the cell cycle. Mutat Res 600(1–2):177–183
Dennison PM, Ramsdale M, Manson CL, Brown AJ (2005) Gene disruption in Candida albicans using a synthetic, codon-optimised Cre-loxP system. Fungal Genet Biol 42(9):737–748
Enloe B, Diamond A, Mitchell AP (2000) A single-transformation gene function test in diploid Candida albicans. J Bacteriol 182(20):5730–5736
Fonzi WA, Irwin MY (1993) Isogenic strain construction and gene mapping in Candida albicans. Genetics 134(3):717–728
Hashimoto S, Ogura M, Aritomi K, Hoshida H, Nishizawa Y, Akada R (2005) Isolation of auxotrophic mutants of diploid industrial yeast strains after UV mutagenesis. Appl Environ Microbiol 71(1):312–319
Hiraoka M, Watanabe K, Umezu K, Maki H (2000) Spontaneous loss of heterozygosity in diploid Saccharomyces cerevisiae cells. Genetics 156(4):1531–1548
Hull CM, Raisner RM, Johnson AD (2000) Evidence for mating of the “asexual” yeast Candida albicans in a mammalian host. Science 289(5477):307–310
Kelly R, Miller SM, Kurtz MB, Kirsch DR (1987) Directed mutagenesis in Candida albicans: one-step gene disruption to isolate ura3 mutants. Mol Cell Biol 7(1):199–208
Magee BB, Magee PT (2000) Induction of mating in Candida albicans by construction of MTLa and MTLalpha strains. Science 289(5477):310–313
Noble SM, Johnson AD (2005) Strains and strategies for large-scale gene deletion studies of the diploid human fungal pathogen Candida albicans. Eukaryot Cell 4(2):298–309
Odds FC (1994) Candida albicans, the life and times of a pathogenic yeast. J Med Vet Mycol 32(Suppl 1):1–8
Poulter R, Hanrahan V, Jeffery K, Markie D, Shepherd MG, Sullivan PA (1982) Recombination analysis of naturally diploid Candida albicans. J Bacteriol 152(3):969–975
Poulter RT (1987) Natural auxotrophic heterozygosity in Candida albicans. Crit Rev Microbiol 15(1):97–101
Reuss O, Vik A, Kolter R, Morschhauser J (2004) The SAT1 flipper, an optimized tool for gene disruption in Candida albicans. Gene 341:119–127
Sadhu C, Hoekstra D, McEachern MJ, Reed SI, Hicks JB (1992) A G-protein alpha subunit from asexual Candida albicans functions in the mating signal transduction pathway of Saccharomyces cerevisiae and is regulated by the a1-alpha 2 repressor. Mol Cell Biol 12(5):1977–1985
Sanyal K, Baum M, Carbon J (2004) Centromeric DNA sequences in the pathogenic yeast Candida albicans are all different and unique. Proc Natl Acad Sci U S A 101(31):11374–11379
Sherman F (1991) Getting started with yeast, vol. 194. Academic, San Diego, CA
Symington LS (2002) Role of RAD52 epistasis group genes in homologous recombination and double-strand break repair. Microbiol Mol Biol Rev 66(4):630–670
Tsang PW, Cao B, Siu PY, Wang J (1999) Loss of heterozygosity, by mitotic gene conversion and crossing over, causes strain-specific adenine mutants in constitutive diploid Candida albicans. Microbiology 145(Pt 7):1623–1629
Tzung KW, Williams RM, Scherer S, Federspiel N, Jones T, Hansen N, Bivolarevic V, Huizar L, Komp C, Surzycki R, Tamse R, Davis RW, Agabian N (2001) Genomic evidence for a complete sexual cycle in Candida albicans. Proc Natl Acad Sci U S A 98(6):3249–53
Wellington M, Rustchenko E (2005) 5-Fluoro-orotic acid induces chromosome alterations in Candida albicans. Yeast 22(1):57–70
Whelan WL, Magee PT (1981) Natural heterozygosity in Candida albicans. J Bacteriol 145(2):896–903
Whelan WL, Partridge RM, Magee PT (1980) Heterozygosity and segregation in Candida albicans. Mol Gen Genet 180(1):107–113
Wilson RB, Davis D, Enloe BM, Mitchell AP (2000) A recyclable Candida albicans URA3 cassette for PCR product-directed gene disruptions. Yeast 16(1):65–70
Wilson RB, Davis D, Mitchell AP (1999) Rapid hypothesis testing with Candida albicans through gene disruption with short homology regions. J Bacteriol 181(6):1868–74
Acknowledgment
We thank Suzanne M. Noble for the C. albicans strains and plasmids. This study was supported by a grant-in-aid for scientific research from the Ministry of Education, Science, Sports, and Culture of Japan and the New Energy and Industrial Technology Department Organization. Y. Takagi is supported by the Twenty-first Century COE Program of the Ministry of Education, Culture, Sports, Science, and Technology.
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Takagi, Y., Akada, R., Kumagai, H. et al. Loss of heterozygosity is induced in Candida albicans by ultraviolet irradiation. Appl Microbiol Biotechnol 77, 1073–1082 (2008). https://doi.org/10.1007/s00253-007-1252-x
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DOI: https://doi.org/10.1007/s00253-007-1252-x