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Molecular and Cellular Biology, December 2008, p. 7504-7513, Vol. 28, No. 24
0270-7306/08/$08.00+0     doi:10.1128/MCB.00137-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Complementary Roles of Yeast Rad4p and Rad34p in Nucleotide Excision Repair of Active and Inactive rRNA Gene Chromatin{triangledown}

Maxime Tremblay,1,{dagger} Yumin Teng,2,{dagger} Michel Paquette,1 Raymond Waters,2 and Antonio Conconi1*

Département de Microbiologie et d'Infectiologie, Faculté de Médecine, Université de Sherbrooke, Sherbrooke, Quebec J1H 5N4, Canada,1 Department of Pathology, School of Medicine, Cardiff University, Health Park, Cardiff CF14 4XN, United Kingdom2

Received 24 January 2008/ Returned for modification 26 February 2008/ Accepted 8 October 2008

Nucleotide excision repair (NER) removes a plethora of DNA lesions. It is performed by a large multisubunit protein complex that finds and repairs damaged DNA in different chromatin contexts and nuclear domains. The nucleolus is the most transcriptionally active domain, and in yeast, transcription-coupled NER occurs in RNA polymerase I-transcribed genes (rDNA). Here we have analyzed the roles of two members of the xeroderma pigmentosum group C family of proteins, Rad4p and Rad34p, during NER in the active and inactive rDNA. We report that Rad4p is essential for repair in the intergenic spacer, the inactive rDNA coding region, and for strand-specific repair at the transcription initiation site, whereas Rad34p is not. Rad34p is necessary for transcription-coupled NER that starts about 40 nucleotides downstream of the transcription initiation site of the active rDNA, whereas Rad4p is not. Thus, although Rad4p and Rad34p share sequence homology, their roles in NER in the rDNA locus are almost entirely distinct and complementary. These results provide evidences that transcription-coupled NER and global genome NER participate in the removal of UV-induced DNA lesions from the transcribed strand of active rDNA. Furthermore, nonnucleosome rDNA is repaired faster than nucleosome rDNA, indicating that an open chromatin structure facilitates NER in vivo.

* Corresponding author. Mailing address: Département de Microbiologie et Infectiologie, Faculté de Médecine, Poste 7446, Université de Sherbrooke, 3001 12th Ave. Nord, Sherbrooke, QC J1H 5N4, Canada. Phone: (819) 564-5360. Fax: (819) 564-5392. E-mail: antonio.conconi{at}usherbrooke.ca

{triangledown} Published ahead of print on 20 October 2008.

{dagger} These authors contributed equally.

Molecular and Cellular Biology, December 2008, p. 7504-7513, Vol. 28, No. 24
0270-7306/08/$08.00+0     doi:10.1128/MCB.00137-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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