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Papers In Press, published online ahead of print January 22, 2002
J. Biol. Chem, 10.1074/jbc.M110941200
Submitted on November 15, 2001
Revised on January 22, 2002
Accepted on January 21, 2002
ETH-Zuerich, Zuerich 8093
Corresponding Author: thoma{at}cell.biol.ethz.ch
DNA-repair by photolyase (photoreactivation) and nucleotide excision repair (NER) are the major pathways to remove UV induced cyclobutane-pyrimidine-dimers (CPDs). The nucleolus is a nuclear subcompartment containing the ribosomal RNA genes (rDNA) of which a fraction is transcribed by RNA polymeraseI (RNAP-I) and the rest is silenced. Here yeast was used to investigate how photoreactivation and NER contribute to repair of active and inactive rDNA. Cells were irradiated with UV light and exposed to different repair conditions. Nuclei were isolated, the active genes were separated from the inactive genes by restriction endonuclease digestion. CPDs were measured in total rDNA, in both fractions, and in the GAL10 gene. Repair in rDNA was as efficient as in GAL10 indicating that both pathways have unrestricted access to the nucleolus. Photoreactivation was much faster than NER and therefore the predominant repair pathway. Active genes were faster repaired by photolyase than silenced genes providing evidence for an open chromatin structure during repair. The transcribed strands of active genes, but not of inactive genes, were slightly faster repaired by NER providing evidence for transcription coupled repair by RNAP-I. There was no pronounced inhibition of photoreactivation by RNAP-I in the transcribed strand, which is in contrast to genes transcribed by RNAP-II and suggests different stabilities of RNAP-I and RNAP-II stalled at CPDs.
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