Dual loss of human POLQ and LIG4 abolishes random integration.
Dual loss of human POLQ and LIG4 abolishes random integration.
复制标题
DOI：
10.1038/ncomms16112
复制
发表时间：
2017-07-11
影响因子：
16.6
通讯作者：
Adachi N
中科院分区：
文献类型：
--
作者：
Saito S;Maeda R;Adachi N
同源重组介导的基因靶向在广泛的物种中极大地促进了遗传分析，但是在人类细胞中是非常低效的，因为极其频繁的随机整合事件，其分子机制仍然是难以捉摸的。在这里，我们表明，DNA聚合酶θ，尽管它在染色体DNA修复中的作用很小，但在很大程度上有助于随机整合，并且缺乏DNA聚合酶θ和DNA连接酶IV的细胞，这是非同源末端连接（NHEJ）所必需的，由于不可检测的随机整合水平，表现出100%的自发基因靶向效率。因此，DNA聚合酶θ介导的末端连接是在不存在NHEJ的情况下唯一的同源性非依赖性修复途径，有趣的是，它们的组合缺失揭示了利用同源性延伸的罕见的β-Alu重组事件。我们的发现为外源DNA整合的机制和DNA聚合酶θ在人类基因组维持中的作用提供了新的见解。由于随机整合事件，同源重组介导的基因靶向在人类细胞中是非常低效的。在这里，作者表明聚合酶θ和DNA连接酶IV的双重抑制消除了随机整合事件。
Homologous recombination-mediated gene targeting has greatly contributed to genetic analysis in a wide range of species, but is highly inefficient in human cells because of overwhelmingly frequent random integration events, whose molecular mechanism remains elusive. Here we show that DNA polymerase θ, despite its minor role in chromosomal DNA repair, substantially contributes to random integration, and that cells lacking both DNA polymerase θ and DNA ligase IV, which is essential for non-homologous end joining (NHEJ), exhibit 100% efficiency of spontaneous gene targeting by virtue of undetectable levels of random integration. Thus, DNA polymerase θ-mediated end joining is the sole homology-independent repair route in the absence of NHEJ and, intriguingly, their combined absence reveals rare Alu-Alu recombination events utilizing a stretch of homology. Our findings provide new insights into the mechanics of foreign DNA integration and the role of DNA polymerase θ in human genome maintenance. Homologous recombination mediated gene targeting is highly inefficient in human cells due to random integration events, Here the authors show that dual repression of polymerase θ and DNA ligase IV eliminate random integration events.
登录
查看更多内容
影响因子：
19
作者：
Ceccaldi R;Rondinelli B;D'Andrea AD
通讯作者：
D'Andrea AD
影响因子：
14.9
作者：
Ahrabi S;Sarkar S;Pfister SX;Pirovano G;Higgins GS;Porter AC;Humphrey TC
通讯作者：
Humphrey TC
影响因子：
16.8
作者：
Kent, Tatiana;Chandramouly, Gurushankar;McDevitt, Shane Michael;Ozdemir, Ahmet Y.;Pomerantz, Richard T.
通讯作者：
Pomerantz, Richard T.
影响因子：
3.5
作者：
Gu, Shen;Yuan, Bo;Lupski, James R.
通讯作者：
Lupski, James R.
影响因子：
14.9
作者：
Arana, Mercedes E.;Seki, Mineaki;Wood, Richard D.;Rogozin, Igor B.;Kunkel, Thomas A.
通讯作者：
Kunkel, Thomas A.