Overexpression of OsRecQl4 and/or OsExo1 Enhances DSB-Induced Homologous Recombination in Rice
Overexpression of OsRecQl4 and/or OsExo1 Enhances DSB-Induced Homologous Recombination in Rice
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DOI：
10.1093/pcp/pcs155
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发表时间：
2012-12-01
影响因子：
4.9
通讯作者：
Toki, Seiichi
中科院分区：
文献类型：
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作者：
Kwon, Yong-Ik;Abe, Kiyomi;Toki, Seiichi
在真核生物的同源重组(HR)介导的DNA双链断裂(DSB)修复过程中，第一步是通过切除5‘端来产生3’-单链DNA(SsDNA)。RAD51在此单链DNA上聚合以搜索同源序列，然后使用未受损的同源DNA链作为模板修复缺口序列。最近在真核生物中的研究表明，DSB位点的切除是由RecQ解旋酶家族蛋白：哺乳动物中的Bloom Helicase(BLM)或酵母中的SGS1和外切核酸酶1(Exo1)共同作用的结果。然而，RecQ解旋酶和核酸外切酶在植物细胞HR 5‘-切除过程中的作用尚不清楚。在这里，我们证明了水稻蛋白OsRecQl4(BLM对应物)和/或OsExo1(Exo1同源物)的过表达可以促进DSB的加工，这是通过水稻重组底物报告系进行的评估。这些结果可用于构建高效的水稻基因打靶系统。
During homologous recombination (HR)-mediated DNA double-strand break (DSB) repair in eukaryotes, an initial step is the creation of a 3'-single-stranded DNA (ssDNA) overhang via resection of a 5' end. Rad51 polymerizes on this ssDNA to search for a homologous sequence, and the gapped sequence is then repaired using an undamaged homologous DNA strand as template. Recent studies in eukaryotes indicate that resection of the DSB site is promoted by the cooperative action of RecQ helicase family proteins: Bloom helicase (BLM) in mammals or Sgs1 in yeast, and exonuclease 1 (Exo1). However, the role of RecQ helicase and exonuclease during the 5'-resection process of HR in plant cells has not yet been defined. Here, we demonstrate that overexpression of rice proteins OsRecQl4 (BLM counterpart) and/or OsExo1 (Exo1 homolog) can enhance DSB processing, as evaluated by recombination substrate reporter lines in rice. These results could be applied to construct an efficient gene targeting system in rice.