Classical non-homologous end-joining pathway utilizes nascent RNA for error-free double-strand break repair of transcribed genes.
Classical non-homologous end-joining pathway utilizes nascent RNA for error-free double-strand break repair of transcribed genes.
复制标题
DOI：
10.1038/ncomms13049
复制
发表时间：
2016-10-05
影响因子：
16.6
通讯作者：
Hazra, Tapas K.
中科院分区：
文献类型：
--
作者：
Chakraborty, Anirban;Tapryal, Nisha;Venkova, Tatiana;Horikoshi, Nobuo;Pandita, Raj K.;Sarker, Altaf H.;Sarkar, Partha S.;Pandita, Tej K.;Hazra, Tapas K.
DNA双链断裂(DSB)导致转录区域的核苷酸丢失可能是致命的。经典非同源末端连接(C-NHEJ)是成年哺乳动物细胞DSB修复(DSBR)的主要途径。在此，我们报道了在这种DSBR过程中，哺乳动物的C-NHEJ蛋白与RNA聚合酶II形成多蛋白复合体，并在DSB诱导后优先与转录基因结合。C-NHEJ因子的缺失显著地消除了转录基因中的DSBR，但在非转录基因中却没有。我们假设新生RNA可以作为恢复缺失序列的模板，从而允许无错误的DSBR。我们确实在C-NHEJ复合体中发现了前mRNA。最后，当一个含有DSB的质粒在E.coliLacZ基因中缺失了几个核苷酸后，在表达LacZ的哺乳动物细胞中被允许有时间修复，一个功能性的LacZ质粒可以从对照细胞中恢复出来，但不能从C-NHEJ因子缺失的细胞中恢复，这为C-NHEJ介导的转录基因组的无错误DSBR提供了重要的机制见解。大多数成年哺乳动物细胞喜欢通过经典的非同源末端连接途径修复双链DNA断裂。在这里，作者提出了新的RNA转录本可以作为模板来促进无错误修复的证据。
DNA double-strand breaks (DSBs) leading to loss of nucleotides in the transcribed region can be lethal. Classical non-homologous end-joining (C-NHEJ) is the dominant pathway for DSB repair (DSBR) in adult mammalian cells. Here we report that during such DSBR, mammalian C-NHEJ proteins form a multiprotein complex with RNA polymerase II and preferentially associate with the transcribed genes after DSB induction. Depletion of C-NHEJ factors significantly abrogates DSBR in transcribed but not in non-transcribed genes. We hypothesized that nascent RNA can serve as a template for restoring the missing sequences, thus allowing error-free DSBR. We indeed found pre-mRNA in the C-NHEJ complex. Finally, when a DSB-containing plasmid with several nucleotides deleted within the E. coli lacZ gene was allowed time to repair in lacZ-expressing mammalian cells, a functional lacZ plasmid could be recovered from control but not C-NHEJ factor-depleted cells, providing important mechanistic insights into C-NHEJ-mediated error-free DSBR of the transcribed genome. Most adult mammalian cells prefer to repair double-strand DNA breaks though the classical nonhomologous end-joining pathway. Here the authors present evidence that a nascent RNA transcript can serve as a template to facilitate error-free repair.
登录
查看更多内容
影响因子：
64.8
作者：
Keskin H;Shen Y;Huang F;Patel M;Yang T;Ashley K;Mazin AV;Storici F
通讯作者：
Storici F
影响因子：
4.8
作者：
Della-Maria, Julie;Zhou, Yi;Tomkinson, Alan E.
通讯作者：
Tomkinson, Alan E.
影响因子：
4.8
作者：
Chakraborty, Anirban;Wakamiya, Maki;Hazra, Tapas K.
通讯作者：
Hazra, Tapas K.
影响因子：
16.8
作者：
Aymard F;Bugler B;Schmidt CK;Guillou E;Caron P;Briois S;Iacovoni JS;Daburon V;Miller KM;Jackson SP;Legube G
通讯作者：
Legube G
影响因子：
64.8
作者：
Francia, Sofia;Michelini, Flavia;Saxena, Alka;Tang, Dave;de Hoon, Michiel;Anelli, Viviana;Mione, Marina;Carninci, Piero;di Fagagna, Fabrizio d'Adda
通讯作者：
di Fagagna, Fabrizio d'Adda