Variations in transcription-repair coupling in mouse cells.
Formation and repair of UV-induced cyclobutane pyrimidine dimers (CPDs) was examined in three different genes in mouse L cells: 1) a stably integrated insert (called LTL), consisting of a herpes simplex virus thymidine kinase gene (tk) fused to a hormone inducible promotor (LTR); 2) the constitutively expressed proto-oncogene c-abl; and 3) the inactive immunoglobulin J chain gene. Transcription of the tk gene is induced > 50-fold by dexamethasone. There is a nonuniform distribution of CPDs in LTL DNA irradiated in vitro, being 4-fold higher in the LTR than in the tk gene, indicating the LTR may be damaged preferentially in irradiated cells. Repair of CPDs occurs efficiently in both strands of LTL and is unaffected by hormone induction of tk gene transcription. Transcription of tk mRNA is very sensitive to UV damage and follows single hit kinetics with UV dose. Furthermore, tk mRNA expression rapidly recovers during repair incubation. Transcription-coupled repair occurs in these cells, however, since only the transcribed strand of c-abl is efficiently repaired of CPDs; the non-transcribed strand as well as both strands of the J chain gene are inefficiently repaired. Thus, repair in the LTL construct may reflect a lack of transcription-coupled repair in either the LTR promotor or the LTL insertion region of chromatin.
Murad
A O
AO
Department of Biochemistry and Biophysics, Washington State University, Pullman 99164-4660.
de Cock
J
J
Brown
D
D
Smerdon
M J
MJ
D000818:Animals; D002460:Cell Line; D004260:DNA Repair; D004251:DNA Transposable Elements; D003907:Dexamethasone / Q000494:pharmacology; D007739:L Cells; D051379:Mice; D011401:Promoter Regions, Genetic; D011740:Pyrimidine Dimers; D012333:RNA, Messenger / Q000096:biosynthesis; D012091:Repetitive Sequences, Nucleic Acid; D013937:Thymidine Kinase / Q000235:genetics; D014158:Transcription, Genetic / Q000528:radiation effects; D014466:Ultraviolet Rays
D004251:DNA Transposable Elements; D011740:Pyrimidine Dimers; D012333:RNA, Messenger; D003907:Dexamethasone; D013937:Thymidine Kinase