Centro de Investigación en Nutrición y Salud Pública, Universidad Autónoma de Nuevo León, Monterrey, NL, México.
FEMS Yeast Res. 2013 Aug;13(5):441-52. doi: 10.1111/1567-1364.12047. Epub 2013 May 2.
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-Δ 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.
解脂耶氏酵母(Yarrowia lipolytica)是一种二相真菌,已成为许多生物学过程的成熟模型,包括异源和嵌合蛋白的分泌。然而,对于负责将编码这些蛋白质的外源 DNA 整合到基因组中的重组机制知之甚少。我们对解脂耶氏酵母的 rad52 缺失突变体进行了表型分析。YlRad52 与其他真核生物(包括酿酒酵母和白色念珠菌)的 Rad52 同源物具有 20-30%的同一性。Ylrad52-Δ 菌株在 YPD-琼脂平板上形成的菌落比野生型的更尖和更小,而在 YPD 液体培养物中,它们的生长速度降低,细胞形态异常,染色质碎片化,这支持同源重组(HR)在非损伤条件下对基因组稳定性的作用。此外,Ylrad52 突变体对紫外线、氧化剂和引起单链(SSB)和双链断裂(DSB)的化合物表现出中度到高度的敏感性,表明 Rad52 在 DNA 修复中起着重要作用。这些发现扩展了 Yl 的先前观察结果,表明 RAD52 是包括酿酒酵母、白色念珠菌和裂殖酵母在内的其他真菌中 DNA 修复的关键基因。