The DNA binding domain and the C-terminal region of DNA Ligase IV specify its role in V(D)J recombination.
The DNA binding domain and the C-terminal region of DNA Ligase IV specify its role in V(D)J recombination.
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DOI：
10.1371/journal.pone.0282236
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发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
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DNA连接酶IV负责修复DNA双链断裂（DSB），包括V（D）J重组过程中产生的DSB。与其他DNA连接酶一样，连接酶IV包含具有三个亚结构域的催化核心-DNA结合（DBD）、核苷酸转移酶（NTD）和寡核苷酸/寡糖折叠亚结构域（OBD）。连接酶IV还具有独特的C-末端区域，其包括两个BRCT结构域、核定位信号序列和参与其与XRCC 4相互作用的一段氨基酸。在三种哺乳动物连接酶中，连接酶IV是参与V（D）J重组并且是V（D）J重组所需的唯一连接酶。在DNA连接酶IV中有助于V（D）J重组的最小结构域的鉴定仍然没有解决。连接酶IV DNA结合结构域与Artemis的相互作用，以及其C-末端区域与XRCC 4的相互作用，表明也与Ku 70/80异二聚体相互作用的这两个区域都很重要，并且可能足以介导DNA连接酶IV参与V（D）J重组。通过交换结构域产生嵌合连接酶蛋白，并测试它们在连接酶IV缺陷细胞中拯救V（D）J重组的能力，研究了这一假设。我们证明了含有连接酶I NTD和两侧为DNA连接酶IV DBD和C-末端区域的OBD的融合蛋白足以支持V（D）J重组。这种嵌合蛋白，我们命名为连接酶37，补充了编码和信号关节的形成。用连接酶37产生的编码接头比用野生型DNA连接酶IV观察到的那些短。较短的长度是由于增加的核苷酸缺失和减少的核苷酸插入。此外，小鼠pro-B细胞系中连接酶37的过表达支持向更短编码接头的转变。我们的研究结果表明，DNA连接酶IV参与V（D）J重组的能力在很大程度上是由其DBD和C-末端区域介导的。
DNA Ligase IV is responsible for the repair of DNA double-strand breaks (DSB), including DSBs that are generated during V(D)J recombination. Like other DNA ligases, Ligase IV contains a catalytic core with three subdomains—the DNA binding (DBD), the nucleotidyltransferase (NTD), and the oligonucleotide/oligosaccharide-fold subdomain (OBD). Ligase IV also has a unique C-terminal region that includes two BRCT domains, a nuclear localization signal sequence and a stretch of amino acid that participate in its interaction with XRCC4. Out of the three mammalian ligases, Ligase IV is the only ligase that participates in and is required for V(D)J recombination. Identification of the minimal domains within DNA Ligase IV that contribute to V(D)J recombination has remained unresolved. The interaction of the Ligase IV DNA binding domain with Artemis, and the interaction of its C-terminal region with XRCC4, suggest that both of these regions that also interact with the Ku70/80 heterodimer are important and might be sufficient for mediating participation of DNA Ligase IV in V(D)J recombination. This hypothesis was investigated by generating chimeric ligase proteins by swapping domains, and testing their ability to rescue V(D)J recombination in Ligase IV-deficient cells. We demonstrate that a fusion protein containing Ligase I NTD and OBDs flanked by DNA Ligase IV DBD and C-terminal region is sufficient to support V(D)J recombination. This chimeric protein, which we named Ligase 37, complemented formation of coding and signal joints. Coding joints generated with Ligase 37 were shorter than those observed with wild type DNA Ligase IV. The shorter length was due to increased nucleotide deletions and decreased nucleotide insertions. Additionally, overexpression of Ligase 37 in a mouse pro-B cell line supported a shift towards shorter coding joints. Our findings demonstrate that the ability of DNA Ligase IV to participate in V(D)J recombination is in large part mediated by its DBD and C-terminal region.
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影响因子：
16
作者：
Grawunder, U;Zimmer, D;Lieber, MR
通讯作者：
Lieber, MR
影响因子：
15.9
作者：
Couedel, Chrystelle;Roman, Christopher;Cortes, Patricia
通讯作者：
Cortes, Patricia
DOI：
10.1084/jem.20111437
发表时间：
2012-05-07
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Malu S;De Ioannes P;Kozlov M;Greene M;Francis D;Hanna M;Pena J;Escalante CR;Kurosawa A;Erdjument-Bromage H;Tempst P;Adachi N;Vezzoni P;Villa A;Aggarwal AK;Cortes P
通讯作者：
Cortes P
影响因子：
10.5
作者：
LIEBER, MR;HESSE, JE;GELLERT, M
通讯作者：
GELLERT, M
DOI：
10.1073/pnas.95.26.15559
发表时间：
1998-12-22
影响因子：
11.1
作者：
Bogue, MA;Jhappan, C;Roth, DB
通讯作者：
Roth, DB