Gene-specific and strand-specific DNA repair in the G1 and G2 phases of the cell cycle.
We have analyzed the fine structure of DNA repair in Chinese hamster ovary (CHO) cells within the G1 and G2 phases of the cell cycle. Repair of inactive regions of the genome has been suggested to increase in the G2 phase of the cell cycle compared with other phases. However, detailed studies of DNA repair in the G2 phase of the cell cycle have been hampered by technical limitations. We have used a novel synchronization protocol (D. K. Orren, L. N. Petersen, and V. A. Bohr, Mol. Cell. Biol. 15:3722-3730, 1995) which permitted detailed studies of the fine structure of DNA repair in G2. CHO cells were synchronized and UV irradiated in G1 or early G2. The rate and extent of removal of cyclobutane pyrimidine dimers from an inactive region of the genome and from both strands of the actively transcribed dihydrofolate reductase (DHFR) gene were examined within each phase. The repair of the transcribed strand of the DHFR gene was efficient in both G1 and G2, with no major differences between the two cell cycle phases. Neither the nontranscribed strand of the DHFR gene nor an inactive region of the genome was repaired in G1 or G2. CHO cells irradiated early in G2 were more resistant to UV irradiation than cells irradiated in late G1. Since we found no major difference in repair rates in G1 and G2, we suggest that G2 resistance can be attributed to the increased time (G2 and G1) available for repair before cells commit to DNA synthesis.
Petersen
L N
LN
Laboratory of Molecular Genetics, National Institute of Aging, Baltimore, Maryland 21224, USA.
Orren
D K
DK
Bohr
V A
VA
D000818:Animals; D016466:CHO Cells; D002453:Cell Cycle / Q000528:radiation effects; D006224:Cricetinae; D004249:DNA Damage; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D005434:Flow Cytometry; D016193:G1 Phase / Q000235:genetics*; D016195:G2 Phase / Q000235:genetics*; D011740:Pyrimidine Dimers / Q000378:metabolism*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*; D013997:Time Factors; D014466:Ultraviolet Rays
D011740:Pyrimidine Dimers; D013762:Tetrahydrofolate Dehydrogenase