Dziadkowiec Dorota, Petters Edyta, Dyjankiewicz Agnieszka, Karpiński Paweł, Garcia Valerie, Watson Adam, Carr Antony M
Faculty of Biotechnology, Wrocław University, Przybyszewskiego 63-77, 51-148 Wrocław, Poland.
DNA Repair (Amst). 2009 May 1;8(5):627-36. doi: 10.1016/j.dnarep.2008.12.008. Epub 2009 Jan 29.
We identified two predicted proteins in Schizosaccharomyces pombe, Rrp1 (SPAC17A2.12) and Rrp2 (SPBC23E6.02) that share 34% and 36% similarity to Saccharomyces cerevisiae Ris1p, respectively. Ris1p is a DNA-dependent ATP-ase involved in gene silencing and DNA repair. Rrp1 and Rrp2 also share similarity with S. cerevisiae Rad5 and S. pombe Rad8, containing SNF2-N, RING finger and Helicase-C domains. To investigate the function of the Rrp proteins, we studied the DNA damage sensitivities and genetic interactions of null mutants with known DNA repair mutants. Single Deltarrp1 and Deltarrp2 mutants were not sensitive to CPT, 4NQO, CDPP, MMS, HU, UV or IR. The double mutants Deltarrp1 Deltarhp51 and Deltarrp2 Deltarhp51 plus the triple Deltarrp1 Deltarrp2 Deltarhp51 mutant did not display significant additional sensitivity. However, the double mutants Deltarrp1 Deltarhp57 and Deltarrp2 Deltarhp57 were significantly more sensitive to MMS, CPT, HU and IR than the Deltarhp57 single mutant. The checkpoint response in these strains was functional. In S. pombe, Rhp55/57 acts in parallel with a second mediator complex, Swi5/Sfr1, to facilitate Rhp51-dependent DNA repair. Deltarrp1 Deltasfr1 and Deltarrp2 Deltasfr1 double mutants did not show significant additional sensitivity, suggesting a function for Rrp proteins in the Swi5/Sfr1 pathway of DSB repair. Consistent with this, Deltarrp1 Deltarhp57 and Deltarrp2 Deltarhp57 mutants, but not Deltarrp1 Deltasfr1 or Deltarrp2 Deltasfr1 double mutants, exhibited slow growth and aberrations in cell and nuclear morphology that are typical of Deltarhp51.
我们在粟酒裂殖酵母中鉴定出两种预测蛋白,即Rrp1(SPAC17A2.12)和Rrp2(SPBC23E6.02),它们与酿酒酵母Ris1p的相似性分别为34%和36%。Ris1p是一种参与基因沉默和DNA修复的依赖DNA的ATP酶。Rrp1和Rrp2也与酿酒酵母Rad5和粟酒裂殖酵母Rad8具有相似性,包含SNF2-N、RING指结构域和解旋酶-C结构域。为了研究Rrp蛋白的功能,我们研究了缺失突变体与已知DNA修复突变体的DNA损伤敏感性和遗传相互作用。单个Deltarrp1和Deltarrp2突变体对喜树碱、4-硝基喹啉-1-氧化物、顺二氯二氨铂、甲基磺酸甲酯、羟基脲、紫外线或电离辐射不敏感。双突变体Deltarrp1 Deltarhp51和Deltarrp2 Deltarhp51以及三突变体Deltarrp1 Deltarrp2 Deltarhp51均未表现出明显的额外敏感性。然而,双突变体Deltarrp1 Deltarhp57和Deltarrp2 Deltarhp57对甲基磺酸甲酯、喜树碱、羟基脲和电离辐射的敏感性明显高于Deltarhp57单突变体。这些菌株中的检查点反应是功能性的。在粟酒裂殖酵母中,Rhp55/57与第二种介导复合物Swi5/Sfr1平行发挥作用,以促进依赖Rhp51的DNA修复。Deltarrp1 Deltasfr1和Deltarrp2 Deltasfr1双突变体未表现出明显的额外敏感性,这表明Rrp蛋白在双链断裂修复的Swi5/Sfr1途径中发挥作用。与此一致的是,Deltarrp1 Deltarhp57和Deltarrp2 Deltarhp57突变体,但不是Deltarrp1 Deltasfr1或Deltarrp2 Deltasfr1双突变体,表现出生长缓慢以及细胞和细胞核形态异常,这是Deltarhp51的典型特征。