We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Repair rates of R-band, G-band and C-band DNA in murine and human cultured cells.
- Authors
Sanders, M. H.; Bates, S. E.; Wilbur, B. S.; Holmquist, G. P.
- Abstract
Repair of cyclobutane pyrimidine dimers (CPDs) in cultured neonatal human fibroblasts and in Mus spretus × M. castaneus F1 neonatal skin fibroblasts was analyzed after UVC-irradiation by cleavage with T4 endonuclease V cyclopyrimidine dimer glycosylase, alkaline-agarose gel electrophoresis, and Southern blotting. The blots were sequentially probed with 32P-labeled Alu, or B2, to preferentially illuminate R-band DNA, by L1 to preferentially illuminate G-band DNA, and by satellite DNA to illuminate C-band DNA. These three different DNA populations showed slightly different global nucleotide excision repair rates that are in the order of speed, R-band DNA > G-band DNA > C-band DNA. Fibroblasts from out-bred neonatal mice and humans showed similar band-specific repair rate ratios and the global repair rate of murine fibroblasts was almost as rapid as that of the human fibroblasts. The mass distribution of the human Alu-probed signal was further analyzed. Gel mobility data was fitted to a logistic equation to include all Mr values. Hypothetical distributions of DNA randomly cleaved to a particular number-average molecular weight were fit to the logistic gel mobility function to determine how such a randomly cleaved distribution of a particular cleavage frequency would be displayed along the experimental gel. This revealed a rapidly repaired kinetic fraction that represented 17% of the Alu-probed signal (R-band DNA), almost none of the L1 probed signal (G-band DNA), and reflects transcription coupled repair of active genes. The remaining Alu-probed DNA showed a random distribution of UVC-induced CPDs throughout all stages of global nucleotide excision repair. The Alu-probed CPDs disappeared with an excellent fit to first order kinetics and with a half-life of seven hours. Copyright © 2003 S. Karger AG, Basel
- Subjects
DNA; DEOXYRIBOSE; NUCLEIC acids; MARMOSA; CELL culture; CYTOLOGICAL techniques; CULTURES (Biology)
- Publication
Cytogenetic & Genome Research, 2004, Vol 104, Issue 1-4, p35
- ISSN
1424-8581
- Publication type
Article
- DOI
10.1159/000077464