CO-CORRECTION OF THE ERCC1, ERCC4 AND XERODERMA-PIGMENTOSUM GROUP-F DNA-REPAIR DEFECTS IN-VITRO
CO-CORRECTION OF THE ERCC1, ERCC4 AND XERODERMA-PIGMENTOSUM GROUP-F DNA-REPAIR DEFECTS IN-VITRO
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DOI：
10.1002/j.1460-2075.1993.tb06043.x
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发表时间：
1993-09-01
期刊：
影响因子：
11.4
通讯作者：
WOOD, RD
中科院分区：
文献类型：
--
作者：
BIGGERSTAFF, M;SZYMKOWSKI, DE;WOOD, RD
哺乳动物ERCC 1编码的多肽是受损DNA的核苷酸切除修复所需的，并且与酿酒酵母RAD 10同源，其在修复和有丝分裂染色体内重组中起作用。代表修复互补组1的啮齿动物细胞具有非功能性ERCC 1。我们报告说，修复紫外线照射的DNA可以重建啮齿动物组1细胞提取物与纠正蛋白质从HeLa细胞。背景修复最小化采用分级啮齿动物细胞提取物补充人类复制蛋白RPA和PCNA。组1-校正活性具有100 kDa的天然分子量并含有33 kDa ERCC 1多肽，以及对来自啮齿动物组4和着色性干皮病组F（XP-F）细胞的提取物的补充活性。第1组、第4组或XP-F细胞的提取物在体外不相互补充，尽管它们补充来自其他组的提取物。通过免疫印迹可检测的ERCC 1的量在组1、组4和XP-F提取物中减少。来自大肠杆菌的重组ERCC 1仅微弱地纠正组1缺陷。这些数据表明，ERCC 1是具有第4组和XP-F校正活性的功能性蛋白质复合物的一部分。后两者可以彼此等同并且类似于酿酒酵母RAD 1。
The mammalian ERCC1-encoded polypeptide is required for nucleotide excision repair of damaged DNA and is homologous to Saccharomyces cerevisiae RAD10, which functions in repair and mitotic intrachromosomal recombination. Rodent cells representing repair complementation group 1 have nonfunctional ERCC1. We report that repair of UV-irradiated DNA can be reconstituted by combining rodent group 1 cell extracts with correcting protein from HeLa cells. Background repair was minimized by employing fractionated rodent celt extracts supplemented with human replication proteins RPA and PCNA. Group 1-correcting activity has a native molecular mass of 100 kDa and contains the 33 kDa ERCC1 polypeptide, as well as complementing activities for extracts from rodent group 4 and xeroderma pigmentosum group F (XP-F) celts. Extracts of group 1, group 4 or XP-F cells do not complement one another in vitro, although they complement extracts from other groups. The amount of ERCC1 detectable by immunoblotting is reduced in group 1, group 4 and XP-F extracts. Recombinant ERCC1 from Escherichia coli only weakly corrected the group 1 defect. The data suggest that ERCC1 is part of a functional protein complex with group 4 and XP-F correcting activities. The latter two may be equivalent to one another and analogous to S.cerevisiae RAD1.