Takagi Yukinobu, Akada Rinji, Kumagai Hidehiko, Yamamoto Kenji, Tamaki Hisanori
Division of Integrated Life Sciences, Graduate School of Biostudies, Kyoto University, Oiwake-cho, Kitashirakawa, Sakyo-ku, Kyoto, 606-8502, Japan.
Appl Microbiol Biotechnol. 2008 Jan;77(5):1073-82. doi: 10.1007/s00253-007-1252-x. Epub 2007 Nov 17.
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.
白色念珠菌是一种人类真菌病原体,因其临床重要性而受到广泛研究。然而,全面的基因分析进展甚微。这是因为该真菌的二倍体和无性特征阻碍了基因破坏。在本研究中,我们发现紫外线(UV)照射以及诱变处理强烈刺激了携带人工构建杂合性的菌株中的杂合性缺失(LOH)。与处于生长稳定期的细胞相比,UV诱导的LOH在对数生长期的细胞中更频繁地发生。在7号染色体上测试的所有位点都观察到了这一点,除了靠近着丝粒的一个位点。白色念珠菌RAD52在酿酒酵母中的直系同源物据报道参与同源重组的DNA修复,结果表明它是UV诱导的LOH所必需的。这些结果表明,UV照射引起的高效LOH可能是白色念珠菌基因分析的一个突出工具。