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不同CHO细胞系核糖体RNA基因中受损DNA的差异修复与复制

Differential repair and replication of damaged DNA in ribosomal RNA genes in different CHO cell lines.

作者信息

Wauthier E L, Hanawalt P C, Vos J M

机构信息

Department of Microbiology and Immunology, School of Medicine, University of North Carolina, Chapel Hill 27599.

出版信息

J Cell Biochem. 1990 Jun;43(2):173-83. doi: 10.1002/jcb.240430208.

Abstract

We studied the repair of psoralen adducts in the pol I-transcribed ribosomal RNA (rRNA) genes of excision repair competent Chinese hamster ovary (CHO) cell lines, their UV sensitive mutant derivatives, and their UV resistant transformants, which express a human excision repair gene. In the parental cell line CHO-AA8, both monoadducts and interstrand crosslinks are removed efficiently from the rRNA genes, whereas neither adduct is removed in the UV sensitive derivative UV5; removal of both adducts is restored in the UV resistant transformant CHO-5T4 carrying the human excision repair gene ERCC-2. In contrast, removal of psoralen adducts from the rRNA genes is not detected in another parental CHO cell line CHO-9, neither in its UV sensitive derivative 43-3B, nor in its UV resistant transformant 83-G5 carrying the human excision repair gene ERCC-1. In contrast to such intergenomic heterogeneity of repair, persistence of psoralen monoadducts during replication of the rRNA genes occurs equally well in all CHO cell lines tested. From these data, we conclude that: 1) the repair efficiency of DNA damage in the rRNA genes varies between established parental CHO cell lines; 2) the repair pathways of intrastrand adducts and interstrand crosslinks in mammalian cells share, at least, one gene product, i.e., the excision repair gene ERCC-2; 3) replicational bypass of psoralen monoadducts at the CHO rRNA locus occurs similarly on both DNA strands.

摘要

我们研究了补骨脂素加合物在具有切除修复能力的中国仓鼠卵巢(CHO)细胞系、其紫外线敏感突变衍生物及其表达人类切除修复基因的紫外线抗性转化体的RNA聚合酶I转录的核糖体RNA(rRNA)基因中的修复情况。在亲本细胞系CHO-AA8中,单加合物和链间交联都能从rRNA基因中有效去除,而在紫外线敏感衍生物UV5中两种加合物均未被去除;在携带人类切除修复基因ERCC-2的紫外线抗性转化体CHO-5T4中,两种加合物的去除得以恢复。相比之下,在另一个亲本CHO细胞系CHO-9及其紫外线敏感衍生物43-3B,以及携带人类切除修复基因ERCC-1的紫外线抗性转化体83-G5中,均未检测到rRNA基因中补骨脂素加合物的去除。与这种修复的基因组间异质性相反,在所有测试的CHO细胞系中,rRNA基因复制过程中补骨脂素单加合物的持续存在情况相同。根据这些数据,我们得出以下结论:1)rRNA基因中DNA损伤的修复效率在已建立的亲本CHO细胞系之间存在差异;2)哺乳动物细胞中链内加合物和链间交联的修复途径至少共享一种基因产物,即切除修复基因ERCC-2;3)在CHO rRNA基因座处,补骨脂素单加合物在两条DNA链上的复制性绕过情况相似。

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