INDUCTION OF A MUTANT PHENOTYPE IN HUMAN REPAIR PROFICIENT CELLS AFTER OVEREXPRESSION OF A MUTATED HUMAN DNA-REPAIR GENE

被引:19
作者
BELT, PBGM
VANOOSTERWIJK, MF
ODIJK, H
HOEIJMAKERS, JHJ
BACKENDORF, C
机构
[1] LEIDEN UNIV,GORLAEUS LABS,DEPT BIOCHEM,MOLEC GENET LAB,POB 9502,2300 AL LEIDEN,NETHERLANDS
[2] ERASMUS UNIV,DEPT CELL BIOL & GENET,3000 DR ROTTERDAM,NETHERLANDS
关键词
D O I
10.1093/nar/19.20.5633
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Antisense and mutated cDNA of the human excision repair gene ERCC-1 were overexpressed in repair proficient HeLa cells by means of an Epstein-Barr-virus derived cDNA expression vector. Whereas antisense RNA did not influence the survival of the transfected cells, a mutated cDNA generating an ERCC-1 protein with two extra amino acids in a conserved region of its C-terminal part resulted in a significant sensitization of the HeLa transfectants to mitomycin C-induced damage. These results suggest that overexpression of the mutated ERCC-1 protein interferes with proper functioning of the excision repair pathway in repair proficient cells and is compatible with a model in which the mutated ERCC-1 protein competes with the wild-type polypeptide for a specific step in the repair process or for occupation of a site in a repair complex. Apparently, this effect is more pronounced for mitomycin C induced crosslink repair than for UV-induced DNA damage.
引用
收藏
页码:5633 / 5637
页数:5
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