Miyazaki Taiga, Tsai Huei-Fung, Bennett John E
Clinical Mycology Section, Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Curr Genet. 2006 Jul;50(1):11-22. doi: 10.1007/s00294-006-0072-3. Epub 2006 Apr 28.
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 Deltakre29 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.
光滑念珠菌KRE29是酿酒酵母KRE29的直系同源基因。酿酒酵母Kre29p已通过亲和纯化被鉴定为Smc5 - Smc6复合物的一个亚基,该复合物是DNA修复和染色体分离所必需的。然而,酿酒酵母KRE29的突变体表型尚未得到很好的表征,其直系同源基因的功能也均未被报道。在此,我们报告了光滑念珠菌kre29缺失突变体的表型特征。光滑念珠菌Kre29p的缺失导致活力下降,即使在正常生长条件下也表现出细胞周期在S期晚期和中期之间停滞,并且还导致质粒丢失率增加,这意味着Kre29p是有丝分裂染色体传递保真度所必需的。该缺失突变体对高温以及包括紫外线、γ射线、4 - 硝基喹啉 - 1 - 氧化物和甲磺酸甲酯在内的DNA损伤剂表现出更高的敏感性,并且这些表型在KRE29回复整合体中得以恢复。与kre29缺失突变体的表型一致,Kre29p - GFP融合蛋白定位于细胞核中。此外,在暴露于4 - 硝基喹啉 - 1 - 氧化物后,Kre29p - GFP变得浓缩并形成明显的焦点。这些结果表明光滑念珠菌Kre29p参与了DNA修复。据我们所知,这是第一篇关于光滑念珠菌中参与DNA修复的细胞蛋白的报道。