Molecular characterization of the human excision repair gene ERCC-1: cDNA cloning and amino acid homology with the yeast DNA repair gene RAD10.
The human excision repair gene ERCC-1 was cloned after DNA mediated gene transfer to the CHO mutant 43-3B, which is sensitive to ultraviolet light and mitomycin-C. We describe the cloning and sequence analysis of the ERCC-1 cDNA and partial characterization of the gene. ERCC-1 has a size of 15 kb and is located on human chromosome 19. The ERCC-1 precursor RNA is subject to alternative splicing of an internal 72 bp coding exon. Only the cDNA of the larger 1.1 kb transcript, encoding a protein of 297 amino acids, was able to confer resistance to ultraviolet light and mitomycin-C on 43-3B cells. Significant amino acid sequence homology was found between the ERCC-1 gene product and the yeast excision repair protein RAD10. The most homologous region displayed structural homology with DNA binding domains of various polypeptides.
M
M
van Duin
J
J
de Wit
H
H
Odijk
A
A
Westerveld
A
A
Yasui
M H
MH
Koken
J H
JH
Hoeijmakers
D
D
Bootsma
D000595:Amino Acid Sequence; D000818:Animals; D001221:Aspartate Carbamoyltransferase; D001483:Base Sequence; D005075:Biological Evolution; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing); D002872:Chromosome Deletion; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000032:analysis*; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000032:analysis; D004080:Dihydroorotase; D005796:Genes; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D011061:Poly A / Q000378:metabolism; D011506:Proteins / Q000032:analysis; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger; D012441:Saccharomyces cerevisiae / Q000235:genetics*
C033626:CAD trifunctional enzyme; D004268:DNA-Binding Proteins; D009097:Multienzyme Complexes; D011506:Proteins; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D004247:DNA; D001221:Aspartate Carbamoyltransferase; D004080:Dihydroorotase; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)