Hope Justin C, Mense Sarah M, Jalakas Merle, Mitsumoto Jun, Freyer Greg A
Graduate Program in Anatomy and Cell Biology, Columbia University, Kolb Building Room 140, 722 West 168th Street, New York, NY 10032, USA.
Proc Natl Acad Sci U S A. 2006 Apr 11;103(15):5875-80. doi: 10.1073/pnas.0601571103. Epub 2006 Apr 4.
Many questions remain about the process of DNA double strand break (DSB) repair by homologous recombination (HR), particularly concerning the exact function played by individual proteins and the details of specific steps in this process. Some recent studies have shown that RecQ DNA helicases have a function in HR. We studied the role of the RecQ helicase Rqh1 with HR proteins in the repair of a DSB created at a unique site within the Schizosaccharomyces pombe genome. We found that DSBs in rqh1(+) cells, are predominantly repaired by interchromosomal gene conversion, with HR between sister chromatids [sister-chromatid conversion (SCC)], occurring less frequently. In Deltarqh1 cells, repair by SCC is favored, and gene conversion rates slow significantly. When we limited the potential for SCC in Deltarqh1 cells by reducing the length of the G2 phase of the cell cycle, DSB repair continued to be predominated by SCC, whereas it was essentially eliminated in wild-type cells. These data indicate that Rqh1 acts to regulate DSB repair by blocking SCC. Interestingly, we found that this role for Rqh1 is independent of its helicase activity. In the course of these studies, we also found nonhomologous end joining to be largely faithful in S. pombe, contrary to current belief. These findings provide insight into the regulation of DSB repair by RecQ helicases.
关于通过同源重组(HR)修复DNA双链断裂(DSB)的过程仍存在许多问题,特别是关于单个蛋白质所起的确切作用以及该过程中特定步骤的细节。最近的一些研究表明,RecQ DNA解旋酶在HR中具有功能。我们研究了RecQ解旋酶Rqh1与HR蛋白在裂殖酵母基因组内一个独特位点产生的DSB修复中的作用。我们发现,rqh1(+)细胞中的DSB主要通过染色体间基因转换进行修复,姐妹染色单体之间的HR[姐妹染色单体转换(SCC)]发生频率较低。在Δrqh1细胞中,SCC修复更受青睐,基因转换率显著减慢。当我们通过缩短细胞周期的G2期长度来限制Δrqh1细胞中SCC的可能性时,DSB修复仍然以SCC为主导,而在野生型细胞中它基本上被消除。这些数据表明,Rqh1通过阻断SCC来调节DSB修复。有趣的是,我们发现Rqh1的这一作用与其解旋酶活性无关。在这些研究过程中,我们还发现非同源末端连接在裂殖酵母中基本是忠实的,这与目前的看法相反。这些发现为RecQ解旋酶对DSB修复的调节提供了见解。