The human CSB (ERCC6) gene corrects the transcription-coupled repair defect in the CHO cell mutant UV61.
The human CSB gene, mutated in Cockayne's syndrome group B (partially defective in both repair and transcription) was previously cloned by virtue of its ability to correct the moderate UV sensitivity of the CHO mutant UV61. To determine whether the defect in UV61 is the hamster equivalent of Cockayne's syndrome, the RNA polymerase II transcription and DNA repair characteristics of a repair-proficient CHO cell line (AA8), UV61 and a CSB transfectant of UV61 were compared. In each cell line, formation and removal of UV-induced cyclobutane pyrimidine dimers (CPDs) were measured in the individual strands of the actively transcribed DHFR gene and in a transcriptionally inactive region downstream of DHFR. AA8 cells efficiently remove CPDs from the transcribed strand, but not from either the non-transcribed strand or the inactive region. There was no detectable repair of CPDs in any region of the genome in UV61. Transfection of the human CSB gene into UV61 restores the normal repair pattern (CPD removal in only the transcribed strand), demonstrating that the DNA repair defect in UV61 is homologous to that in Cockayne's syndrome (complementation group B) cells. However, we observe no significant deficiency in RNA polymerase II-mediated transcription in UV61, suggesting that the CSB protein has independent roles in DNA repair and RNA transcription pathways.
Orren
D K
DK
Laboratory of Molecular Genetics, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA.
Dianov
G L
GL
Bohr
V A
VA
D000818:Animals; D016466:CHO Cells; D002470:Cell Survival; D002474:Cell-Free System; D003057:Cockayne Syndrome / Q000235:genetics*; D006224:Cricetinae / Q000235:genetics*; D004265:DNA Helicases / Q000235:genetics*; D004257:DNA Polymerase II / Q000378:metabolism; D004260:DNA Repair / Q000235:genetics*; D045643:DNA Repair Enzymes; D004307:Dose-Response Relationship, Radiation; D016259:Genes, myc; D005816:Genetic Complementation Test; D006801:Humans; D009154:Mutation; D000075223:Poly-ADP-Ribose Binding Proteins; D011740:Pyrimidine Dimers / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection; D014466:Ultraviolet Rays / Q000009:adverse effects
D000075223:Poly-ADP-Ribose Binding Proteins; D011740:Pyrimidine Dimers; D004257:DNA Polymerase II; D004265:DNA Helicases; C078673:ERCC6 protein, human; D045643:DNA Repair Enzymes