裂殖酵母 Mms1 通道将受干扰的复制修复为 Rhp51 独立的同源重组。
Schizosaccharomyces pombe Mms1 channels repair of perturbed replication into Rhp51 independent homologous recombination.
机构信息
Cell Cycle and Genome Stability Group, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, 2200 Copenhagen N., Denmark.
出版信息
DNA Repair (Amst). 2011 Mar 7;10(3):283-95. doi: 10.1016/j.dnarep.2010.11.013. Epub 2010 Dec 23.
In both Schizosaccharomyces pombe and Saccharomyces cerevisiae, Mms22 and Mms1 form a complex with important functions in the response to DNA damage, loss of which leads to perturbations during replication. Furthermore, in S. cerevisiae, Mms1 has been suggested to function in concert with a Cullin-like protein, Rtt101/Cul8, a potential paralog of Cullin 4. We performed epistasis analysis between Δmms1 and mutants of pathways with known functions in genome integrity, and measured the recruitment of homologous recombination proteins to blocked replication forks and recombination frequencies. We show that, in S. pombe, the functions of Mms1 and the conserved components of the Cullin 4 ubiquitin ligase, Pcu4 and Ddb1, do not significantly overlap. Furthermore, unlike in S. cerevisiae, the function of the H3K56 acetylase Rtt109 is not essential for Mms1 function. We provide evidence that Mms1 function is particularly important when a single strand break is converted into a double strand break during replication. Genetic data connect Mms1 to a Mus81 and Rad22(Rad52) dependent, but Rhp51 independent, branch of homologous recombination. This is supported by results demonstrating that Mms1 is recruited to a site-specific replication fork barrier and that, in a Δmms1 strain, Rad22(Rad52) and RPA recruitment to blocked forks are reduced, whereas Rhp51 recruitment is unaffected. In addition, Mms1 appears to specifically promote chromosomal rearrangements in a recombination assay. These observations suggest that Mms1 acts to channel repair of perturbed replication into a particular sub-pathway of homologous recombination.
在酿酒酵母和裂殖酵母中,Mms22 和 Mms1 形成一个复合物,在应对 DNA 损伤方面具有重要功能,其缺失会导致复制过程中的干扰。此外,在酿酒酵母中,Mms1 被认为与一种 Cullin 样蛋白 Rtt101/Cul8 协同作用,后者是 Cullin 4 的潜在同源物。我们在基因组完整性已知功能的途径突变体与Δmms1 之间进行了上位性分析,并测量了同源重组蛋白在受阻复制叉上的募集和重组频率。我们表明,在裂殖酵母中,Mms1 的功能与 Cullin 4 泛素连接酶的保守成分 Pcu4 和 Ddb1 没有显著重叠。此外,与酿酒酵母不同,H3K56 乙酰转移酶 Rtt109 的功能对于 Mms1 功能不是必需的。我们提供的证据表明,当在复制过程中单个链断裂转化为双链断裂时,Mms1 功能尤为重要。遗传数据将 Mms1 与 Mus81 和 Rad22(Rad52)依赖性但 Rhp51 非依赖性同源重组分支联系起来。这一结果得到了以下结果的支持:证明 Mms1 被募集到特定的复制叉障碍位点,并且在Δmms1 菌株中,Rad22(Rad52)和 RPA 募集到受阻的叉点减少,而 Rhp51 的募集不受影响。此外,Mms1 似乎在重组测定中特别促进染色体重排。这些观察结果表明,Mms1 作用是将受损复制的修复引导到同源重组的特定亚途径中。