Structure and functional analysis of the BRCT domain of translesion synthesis DNA polymerase Rev1.
Structure and functional analysis of the BRCT domain of translesion synthesis DNA polymerase Rev1.
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DOI：
10.1021/bi301572z
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发表时间：
2013-01-08
期刊：
影响因子：
2.9
通讯作者：
Washington MT
中科院分区：
文献类型：
--
作者：
Pryor JM;Gakhar L;Washington MT
转录合成（TLS）是一种使用专门的低保真度DNA聚合酶来克服DNA损伤引起的复制障碍的途径。这种途径的使用通常会导致体细胞突变，从而导致癌变。Rev 1是在所有真核生物中发现的TLS聚合酶，其在介导DNA损伤诱导的诱变中起关键作用。它具有BRCA 1 C-末端（BRCT）结构域，这是其功能所必需的。Rev 1 -1等位基因编码Rev 1的突变形式，在该结构域中具有G193 R取代，其减少DNA损伤诱导的诱变。尽管BRCT结构域在致突变性TLS中具有明显的重要性，但其作用尚不清楚。在这里，我们报告的X-射线晶体结构的酵母Rev 1 BRCT域，并表明在残基构成其磷酸盐结合口袋的取代不影响诱变TLS。这表明Rev 1 BRCT结构域的作用不是识别蛋白质结合伴侣或DNA上的磷酸基团。我们还发现，G193残基位于BRCT结构域的一个保守的转弯区域，我们的体内和体外研究表明，G193 R取代可能会破坏Rev 1的功能，通过破坏BRCT结构域的折叠。
Translesion synthesis (TLS) is a pathway in which specialized, low-fidelity DNA polymerases are used to overcome replication blocks caused by DNA damage. The use of this pathway often results in somatic mutations that can drive carcinogenesis. Rev1 is a TLS polymerase found in all eukaryotes that plays a pivotal role in mediating DNA-damage induced mutagenesis. It possesses a BRCA1 C-terminal (BRCT) domain that is required for its function. The rev1-1 allele encodes a mutant form of Rev1 with a G193R substitution in this domain, which reduces DNA damage-induced mutagenesis. Despite its clear importance in mutagenic TLS, the role of the BRCT domain is unknown. Here, we report the X-ray crystal structure of the yeast Rev1 BRCT domain and show that substitutions in residues constituting its phosphate-binding pocket do not affect mutagenic TLS. This suggests that the role of the Rev1 BRCT domain is not to recognize phosphate groups on protein binding partners or on DNA. We also found that the G193 residue is located in a conserved turn region of the BRCT domain, and our in vivo and in vitro studies suggest that the G193R substitution may disrupt Rev1 function by destabilizing the fold of the BRCT domain.
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DOI：
10.1107/s090744491003982x
发表时间：
2011-04
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Evans PR
通讯作者：
Evans PR
影响因子：
16
作者：
Guo, Caixia;Sonoda, Eiichiro;Friedberg, Errol C.
通讯作者：
Friedberg, Errol C.
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
14.9
作者：
Howell, Craig A.;Kondratick, Christine M.;Washington, M. Todd
通讯作者：
Washington, M. Todd
影响因子：
3.9
作者：
Lawrence, CW;Maher, VM
通讯作者：
Maher, VM