Expression and nucleotide excision repair of a UV-irradiated reporter gene in unirradiated human cells.
It has been suggested that reactivation of damaged reporter genes introduced into cultured mammalian cells reflects transcription-coupled nucleotide excision repair. To evaluate this possibility directly, we introduced a UV-irradiated shuttle vector, pCMV beta, into unirradiated human cells and compared expression of the reporter gene (lacZ) with repair of cyclobutane pyrimidine dimers (CPDs). Expression of the irradiated reporter gene was more UV resistant in XPC cells, which are deficient in global genome repair, than in CSB cells, which are deficient in transcription-coupled repair. These results are consistent with the idea that repair of the reporter gene is primarily dependent upon transcription-coupled repair. However, when the plasmid DNA was analyzed for removal of CPDs, no clear evidence was obtained for transcription-coupled repair either in XPC cells or in cells with normal repair capacity.
Ganesan
A K
AK
Department of Biological Sciences, Stanford University, CA 94305-5020, USA. anngan@leland.stanford.edu
Hunt
J
J
Hanawalt
P C
PC
D002460:Cell Line; D004260:DNA Repair; D017930:Genes, Reporter / Q000528:radiation effects*; D006801:Humans; D007763:Lac Operon; D011740:Pyrimidine Dimers / Q000235:genetics; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics
D011740:Pyrimidine Dimers