Differential Roles for DNA Polymerases Eta, Zeta, and REV1 in Lesion Bypass of Intrastrand versus Interstrand DNA Cross-Links
Differential Roles for DNA Polymerases Eta, Zeta, and REV1 in Lesion Bypass of Intrastrand versus Interstrand DNA Cross-Links
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DOI：
10.1128/mcb.00993-09
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发表时间：
2010-03-01
影响因子：
5.3
通讯作者：
Canman, Christine E.
中科院分区：
文献类型：
--
作者：
Hicks, J. Kevin;Chute, Colleen L.;Canman, Christine E.
转录DNA合成（TLS）是一个过程，其中专门的DNA聚合酶被招募，以绕过DNA病变，否则将停止高保真聚合酶。我们提供的证据表明，跨顺铂链内交联的TLS是由多个translesion DNA聚合酶进行的。首先，我们确定了通过RAD 18的PCNA单泛素化对于通过TLS聚合酶eta（Pol eta）、REV 1和zeta（Pol zeta）有效绕过顺铂加合物是必要的，这是基于这些蛋白质的消耗单独导致细胞存活率降低、S期细胞周期停滞和DNA损伤反应活化的观察结果。第二，我们表明，除了通过RAD 18的PCNA单泛素化之外，范可尼贫血核心复合物对于募集REV 1到顺铂处理的细胞中停滞的复制叉也是重要的。第三，我们提出的证据表明，REV1和Pol zeta是唯一与顺铂和丝裂霉素C诱导的染色体畸变的保护，都是必要的，及时解决DNA双链断裂与修复DNA链间交联。总之，我们的研究结果表明，REV1和Pol zeta促进修复链间交联独立的PCNA monoubiquitination和Pol eta，而RAD 18加上Pol eta，REV1，和Pol zeta都是必要的复制旁路顺铂链内DNA交联。
Translesion DNA synthesis (TLS) is a process whereby specialized DNA polymerases are recruited to bypass DNA lesions that would otherwise stall high-fidelity polymerases. We provide evidence that TLS across cisplatin intrastrand cross-links is performed by multiple translesion DNA polymerases. First, we determined that PCNA monoubiquitination by RAD18 is necessary for efficient bypass of cisplatin adducts by the TLS polymerases eta (Pol eta), REV1, and zeta (Pol zeta) based on the observations that depletion of these proteins individually leads to decreased cell survival, cell cycle arrest in S phase, and activation of the DNA damage response. Second, we showed that in addition to PCNA monoubiquitination by RAD18, the Fanconi anemia core complex is also important for recruitment of REV1 to stalled replication forks in cisplatin treated cells. Third, we present evidence that REV1 and Pol zeta are uniquely associated with protection against cisplatin and mitomycin C-induced chromosomal aberrations, and both are necessary for the timely resolution of DNA double-strand breaks associated with repair of DNA interstrand cross-links. Together, our findings indicate that REV1 and Pol zeta facilitate repair of interstrand cross-links independently of PCNA monoubiquitination and Pol eta, whereas RAD18 plus Pol eta, REV1, and Pol zeta are all necessary for replicative bypass of cisplatin intrastrand DNA cross-links.