Functional capacity of XRCC1 protein variants identified in DNA repair-deficient Chinese hamster ovary cell lines and the human population.
Functional capacity of XRCC1 protein variants identified in DNA repair-deficient Chinese hamster ovary cell lines and the human population.
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DOI：
10.1093/nar/gkq193
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发表时间：
2010-08
影响因子：
14.9
通讯作者：
Wilson DM 3rd
中科院分区：
文献类型：
--
作者：
Berquist BR;Singh DK;Fan J;Kim D;Gillenwater E;Kulkarni A;Bohr VA;Ackerman EJ;Tomkinson AE;Wilson DM 3rd
XRCC 1在碱基切除修复中作为支架蛋白发挥作用，这是一种处理DNA中碱基和糖损伤的途径。利用重组XRCC 1蛋白的研究表明：C389 Y突变导致蛋白质不稳定，V86 R突变导致与POLβ的相互作用消失，但不影响与PARP-1、LIG 3 α和PCNA的相互作用;在EM-C12中鉴定的E98 K取代降低了蛋白质完整性，略微破坏了POLβ相互作用的稳定性，并略微增强了DNA结合。两种罕见的（P161 L和Y 576 S）和两种常见的（R194 W和R399 Q）氨基酸群体变异对XRCC 1蛋白的稳定性或与POLβ、PARP-1、LIG 3 α、PCNA或DNA的相互作用影响很小或没有影响。一种常见的群体变异体（R280 H）对与POLβ、PARP-1、LIG 3 α和PCNA的相互作用没有明显影响，但确实降低了DNA结合能力。当在HeLa细胞中表达时，XRCC 1变体-不包括E98 K（主要是核仁）和C389 Y（表达减少）-表现出正常的核分布。大多数蛋白质变体，包括V86 R POLβ相互作用突变体，显示出正常的再定位动力学，往返于激光诱导的DNA损伤位点：除了E98 K和C389 Y，以及多态性变体R280 H，其在DNA断裂处显示出稍短的保留时间。
XRCC1 operates as a scaffold protein in base excision repair, a pathway that copes with base and sugar damage in DNA. Studies using recombinant XRCC1 proteins revealed that: a C389Y substitution, responsible for the repair defects of the EM-C11 CHO cell line, caused protein instability; a V86R mutation abolished the interaction with POLβ, but did not disrupt the interactions with PARP-1, LIG3α and PCNA; and an E98K substitution, identified in EM-C12, reduced protein integrity, marginally destabilized the POLβ interaction, and slightly enhanced DNA binding. Two rare (P161L and Y576S) and two frequent (R194W and R399Q) amino acid population variants had little or no effect on XRCC1 protein stability or the interactions with POLβ, PARP-1, LIG3α, PCNA or DNA. One common population variant (R280H) had no pronounced effect on the interactions with POLβ, PARP-1, LIG3α and PCNA, but did reduce DNA-binding ability. When expressed in HeLa cells, the XRCC1 variants—excluding E98K, which was largely nucleolar, and C389Y, which exhibited reduced expression—exhibited normal nuclear distribution. Most of the protein variants, including the V86R POLβ-interaction mutant, displayed normal relocalization kinetics to/from sites of laser-induced DNA damage: except for E98K and C389Y, and the polymorphic variant R280H, which exhibited a slightly shorter retention time at DNA breaks.
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影响因子：
14.9
作者：
Fan, JS;Otterlei, M;Wilson, DM
通讯作者：
Wilson, DM
影响因子：
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影响因子：
14.9
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影响因子：
14.9
作者：
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