Fine structural analysis of DNA repair in mammalian cells.
Nucleotide excision repair (NER) of ultraviolet (UV) light induced photo lesions is heterogeneous in the genomic DNA. We have investigated the mechanistic basis for this repair heterogeneity by analyzing NER activity in higher order chromatin of repair proficient hamster cells. Immunological labeling of repair and transcription sites indicates that NER initiates at the nuclear matrix in close association with transcription. The repair gradually extends into the loop DNA regions in a time dependent fashion. Repair analysis indicates that the DNA damaged by UV irradiation is recruited to the nuclear matrix soon after UV exposure. Consistent with this finding, immunofluorescence and western blotting analyses indicate the enrichment of many NER proteins (XPA, RPA, PCNA, the P62 and p89 sub-units of the basal transcription factor, TFIIH) in the nuclear matrix of UV treated cells. These results strengthen the notion that the nuclear matrix is an important site for the assembly of an efficient repair complex.
Balajee
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AS
Laboratory of Molecular Genetics, National Institute on Aging, National Institute of Health, 5600, Nathan Shock Drive, Baltimore, MD 212224, USA. balajeea@grc.nia.nih.gov
May
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Bohr
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VA
D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004249:DNA Damage / Q000528:radiation effects*; D004260:DNA Repair / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D015530:Nuclear Matrix / Q000502:physiology; D009687:Nuclear Proteins / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000528:radiation effects; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics; D014466:Ultraviolet Rays
D002843:Chromatin; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D014157:Transcription Factors; D001973:Bromodeoxyuridine