A multistep genomic screen identifies new genes required for repair of DNA double-strand breaks in Saccharomyces cerevisiae.
A multistep genomic screen identifies new genes required for repair of DNA double-strand breaks in Saccharomyces cerevisiae.
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DOI：
10.1186/1471-2164-14-251
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发表时间：
2013-04-15
期刊：
影响因子：
4.4
通讯作者：
Lewis LK
中科院分区：
文献类型：
--
作者：
McKinney JS;Sethi S;Tripp JD;Nguyen TN;Sanderson BA;Westmoreland JW;Resnick MA;Lewis LK
DNA双链断裂（DSB）的有效重新连接机制是至关重要的，因为这种损伤的错误修复会导致突变，非整倍体和细胞活力丧失。DSB修复是由蛋白质介导的两个主要途径，称为同源重组和非同源末端连接。修复效率也受到其他过程的调节，如姐妹染色单体凝聚力，核小体重塑和DNA损伤检查点。影响DSB修复效率的基因总数是未知的。为了鉴定影响DSB修复的新的酵母基因，测试了在先前的全基因组调查中与γ辐射抗性相关的基因对由EcoRI内切核酸酶的体内表达产生的位点特异性DSB的修复的影响。在利用MAT α和MATα缺失菌株文库的筛选中，发现DNA修复基因RAD52组的8个成员（RAD50、RAD51、RAD52、RAD54、RAD55、RAD57、MRE 11和XRS2）和73个额外的基因是有效修复EcoRI诱导的DSB所需的。大多数突变体也敏感的染色体断裂化学物质MMS和博莱霉素。一些非RAD52组基因先前已与DNA修复有关，超过一半的基因影响核过程。在高通量蛋白质组学研究中，许多由保护性基因编码的蛋白质先前已被证明彼此物理相关，并与已知的DNA修复蛋白质相关联。大多数蛋白质（64%）与人类蛋白质具有序列相似性，表明它们具有相似的功能。我们已经使用了一种遗传筛选方法来检测所需的新基因的有效修复的DSB在酿酒酵母。这些发现突出了对维持基因组完整性至关重要的新基因，因此当相应的基因直系同源物和同源物在人类细胞中失活或多态时，它们的潜在影响最令人担忧。
Efficient mechanisms for rejoining of DNA double-strand breaks (DSBs) are vital because misrepair of such lesions leads to mutation, aneuploidy and loss of cell viability. DSB repair is mediated by proteins acting in two major pathways, called homologous recombination and nonhomologous end-joining. Repair efficiency is also modulated by other processes such as sister chromatid cohesion, nucleosome remodeling and DNA damage checkpoints. The total number of genes influencing DSB repair efficiency is unknown. To identify new yeast genes affecting DSB repair, genes linked to gamma radiation resistance in previous genome-wide surveys were tested for their impact on repair of site-specific DSBs generated by in vivo expression of EcoRI endonuclease. Eight members of the RAD52 group of DNA repair genes (RAD50, RAD51, RAD52, RAD54, RAD55, RAD57, MRE11 and XRS2) and 73 additional genes were found to be required for efficient repair of EcoRI-induced DSBs in screens utilizing both MATa and MATα deletion strain libraries. Most mutants were also sensitive to the clastogenic chemicals MMS and bleomycin. Several of the non-RAD52 group genes have previously been linked to DNA repair and over half of the genes affect nuclear processes. Many proteins encoded by the protective genes have previously been shown to associate physically with each other and with known DNA repair proteins in high-throughput proteomics studies. A majority of the proteins (64%) share sequence similarity with human proteins, suggesting that they serve similar functions. We have used a genetic screening approach to detect new genes required for efficient repair of DSBs in Saccharomyces cerevisiae. The findings have spotlighted new genes that are critical for maintenance of genome integrity and are therefore of greatest concern for their potential impact when the corresponding gene orthologs and homologs are inactivated or polymorphic in human cells.
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影响因子：
56.9
作者：
Hillenmeyer, Maureen E.;Fung, Eula;Giaever, Guri
通讯作者：
Giaever, Guri
DOI：
10.1073/pnas.0909917107
发表时间：
2010-03-02
影响因子：
11.1
作者：
Bahmed, Karim;Nitiss, Karin C.;Nitiss, John L.
通讯作者：
Nitiss, John L.
影响因子：
30.8
作者：
Bennett, CB;Lewis, LK;Resnick, MA
通讯作者：
Resnick, MA
影响因子：
14.9
作者：
Ahne, F;Jha, B;EckardtSchupp, F
通讯作者：
EckardtSchupp, F
影响因子：
3.3
作者：
Game, JC;Williamson, MS;Baccari, C
通讯作者：
Baccari, C