DNA repair of pyrimidine dimers and 6-4 photoproducts in the ribosomal DNA.

AUTOR(ES)

Balajee, A S

RESUMO

The nucleolus is a unique structural component of interphase nuclei where the ribosomal genes, trans-cribed by RNA polymerase I (RNA pol I), are organized. In the present study, the repair of UV-induced photolesions was investigated in the ribosomal DNA (rDNA) in relation to RNA pol I transcription. We used hamster cells because their repair phenotype permits the separate analysis of the major photo-products induced by UV light. Immunofluorescent labeling of UV-induced DNA repair and transcription sites showed that the nucleolar regions were defic-ient in DNA repair despite the presence of abundant RNA pol I transcription foci. Immunological staining indicated that various NER proteins, including TFIIH (subunits p62 and p89), p53, Gadd 45 and prolifer-ating cell nuclear antigen are all enriched in the nuclei but distinctly absent in nucleoli. This lack of enrichment of NER factors in the nucleolus may be responsible for the inefficient repair of photo-products in the rDNA. UV irradiation generates two major photoproducts, the cyclobutane pyrimidine dimers (CPDs) and the 6-4 photoproducts (6-4 PPs). The repair kinetics of these two lesions were assessed simultaneously by the immunological isolation of bromodeoxyuridine (BudR) containing excision repair patches using an antibody to BudR. We found that the repair of the photolesions was less efficient in the rDNA compared to that of the endo-genous housekeeping gene, dihydrofolate reductase (DHFR). Gene specific repair of each of these two photoproducts was then measured separately in the rDNA and in the DHFR gene, which is transcribed by RNA pol II. The removal of CPDs was deficient in the rDNA as compared to the DHFR gene. On the contrary, 6-4 PPs were removed efficiently from the rDNA although somewhat slower than from the DHFR gene. The relatively efficient repair of 6-4 PPs in the rDNA is consistent with the notion that the 6-4 PPs are repaired efficiently in different genomic regions by the global genome repair pathway.

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