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The RAD52 ortholog of Yarrowia lipolytica is essential for nuclear integrity and DNA repair

机译:解脂耶氏酵母的RAD52直系同源物对于核完整性和DNA修复至关重要

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Yarrowia lipolytica (Yl) is a dimorphic fungus that has become a well-established model for a number of biological processes, including secretion of heterologous and chimerical proteins. However, little is known on the recombination machinery responsible for the integration in the genome of the exogenous DNA encoding for those proteins. We have carried out a phenotypic analysis of rad52 deletants of Y.lipolytica. YlRad52 exhibited 20-30% identity with Rad52 homologues of other eukaryotes, including Saccharomyces cerevisiae and Candida albicans. Ylrad52- strains formed colonies on YPD-agar plates which were spinier and smaller than those from wild type, whereas in YPD liquid cultures they exhibited a decreased grow rate and contained cells with aberrant morphology and fragmented chromatin, supporting a role for homologous recombination (HR) in genome stability under nondamaging conditions. In addition, Ylrad52 mutants showed moderate to high sensitivity to UV light, oxidizing agents and compounds that cause single- (SSB) and double-strand breaks (DSB), indicating an important role for Rad52 in DNA repair. These findings extend to Yl previous observations indicating that RAD52 is a crucial gene for DNA repair in other fungi, including S.cerevisiae, C.albicans and Schizosaccharomyces pombe.
机译:解脂耶氏酵母(Ylia lipolytica)(Y1)是一种双态真菌,已成为许多生物学过程(包括异源和嵌合蛋白分泌)的公认模型。然而,对于负责编码那些蛋白质的外源DNA在基因组中整合的重组机制知之甚少。我们已经对解脂耶氏酵母的rad52缺失进行了表型分析。 Y1Rad52与包括酿酒酵母和白色念珠菌在内的其他真核生物的Rad52同源物具有20-30%的同一性。 Ylrad52-菌株在YPD-琼脂平板上形成菌落,比野生型琼脂平板上的细胞更尖锐,更小,而在YPD液体培养物中,它们表现出降低的生长速度,并包含形态异常和染色质碎片的细胞,支持同源重组(HR) )在无损条件下的基因组稳定性。此外,Ylrad52突变体对紫外线,氧化剂和会导致单链断裂(SSB)和双链断裂(DSB)的化合物表现出中等至高的敏感性,表明Rad52在DNA修复中具有重要作用。这些发现扩展到Y1以前的观察结果,表明RAD52是其他真菌,包括酿酒酵母,白色念珠菌和粟酒裂殖酵母中DNA修复的关键基因。

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