Rev1 is a base excision repair enzyme with 5'-deoxyribose phosphate lyase activity.
Rev1 is a base excision repair enzyme with 5'-deoxyribose phosphate lyase activity.
复制标题
DOI：
10.1093/nar/gkw869
复制
发表时间：
2016-12-15
影响因子：
14.9
通讯作者：
Wilson SH
中科院分区：
文献类型：
--
作者：
Prasad R;Poltoratsky V;Hou EW;Wilson SH
Rev 1是DNA聚合酶Y家族的成员，以其脱氧胞苷酰转移酶活性（将dCMP掺入DNA中）及其在跨病变旁路事件中作为其他Y家族聚合酶的支架因子的能力而闻名。Rev 1还参与免疫球蛋白基因体细胞超突变过程中的诱变过程。鉴于突变模式与先前在用碱基切除修复诱导剂处理的小鼠成纤维细胞中观察到的dCMP插入一致，我们质疑Rev 1是否也参与碱基切除修复（BER）。在此，我们发现了一个弱的5′-脱氧核糖磷酸（5′-dRP）裂解酶活性在小鼠Rev 1，并证明该酶可以介导BER在体外。全长Rev 1蛋白及其催化核心结构域在体外支持BER的能力方面是相似的。这两种蛋白质中的dRP裂解酶活性通过Schiff碱中间体的NaBH 4还原和动力学研究来证实。有限的蛋白水解，质谱和缺失分析本地化的dRP裂解酶活性位点的C-末端片段Rev 1的催化核心结构域。这些结果表明，Rev 1可以作为BER中的备用聚合酶，并可能通过这种活性促进艾滋病引发的抗体多样化。
Rev1 is a member of the Y-family of DNA polymerases and is known for its deoxycytidyl transferase activity that incorporates dCMP into DNA and its ability to function as a scaffold factor for other Y-family polymerases in translesion bypass events. Rev1 also is involved in mutagenic processes during somatic hypermutation of immunoglobulin genes. In light of the mutation pattern consistent with dCMP insertion observed earlier in mouse fibroblast cells treated with a base excision repair-inducing agent, we questioned whether Rev1 could also be involved in base excision repair (BER). Here, we uncovered a weak 5′-deoxyribose phosphate (5′-dRP) lyase activity in mouse Rev1 and demonstrated the enzyme can mediate BER in vitro. The full-length Rev1 protein and its catalytic core domain are similar in their ability to support BER in vitro. The dRP lyase activity in both of these proteins was confirmed by NaBH4 reduction of the Schiff base intermediate and kinetics studies. Limited proteolysis, mass spectrometry and deletion analysis localized the dRP lyase active site to the C-terminal segment of Rev1's catalytic core domain. These results suggest that Rev1 could serve as a backup polymerase in BER and could potentially contribute to AID-initiated antibody diversification through this activity.
登录
查看更多内容
影响因子：
16
作者：
Guo, Caixia;Sonoda, Eiichiro;Friedberg, Errol C.
通讯作者：
Friedberg, Errol C.
影响因子：
16
作者：
Haracska, L;Kondratick, CM;Prakash, L
通讯作者：
Prakash, L
影响因子：
3.7
作者：
Akre MK;Starrett GJ;Quist JS;Temiz NA;Carpenter MA;Tutt AN;Grigoriadis A;Harris RS
通讯作者：
Harris RS
影响因子：
2.9
作者：
Fortini, P;Pascucci, B;Dogliotti, E
通讯作者：
Dogliotti, E
影响因子：
3.9
作者：
Lawrence, CW;Maher, VM
通讯作者：
Maher, VM