title: RNA polymerase II bypass of oxidative DNA damage is regulated by transcription elongation factors.
abstract: Oxidative lesions represent the most abundant DNA lesions within the cell. In the present study, we investigated the impact of the oxidative lesions 8-oxoguanine, thymine glycol and 5-hydroxyuracil on RNA polymerase II (RNA pol II) transcription using a well-defined in vitro transcription system. We found that in a purified, reconstituted transcription system, these lesions block elongation by RNA pol II to different extents, depending on the type of lesion. Suggesting the presence of a bypass activity, the block to elongation is alleviated when transcription is carried out in HeLa cell nuclear extracts. By purifying this activity, we discovered that TFIIF could promote elongation through a thymine glycol lesion. The elongation factors Elongin and CSB, but not TFIIS, can also stimulate bypass of thymine glycol lesions, whereas Elongin, CSB and TFIIS can all enhance bypass of an 8-oxoguanine lesion. By increasing the efficiency with which RNA pol II reads through oxidative lesions, elongation factors can contribute to transcriptional mutagenesis, an activity that could have implications for the generation or progression of human diseases.
authors.affiliation: Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch Cedex, CU Strasbourg, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Charlet-Berguerand
authors.forename: Sascha
authors.initials: S
authors.lastname: Feuerhahn
authors.forename: Stephanie E
authors.initials: SE
authors.lastname: Kong
authors.forename: Howard
authors.initials: H
authors.lastname: Ziserman
authors.forename: Joan W
authors.initials: JW
authors.lastname: Conaway
authors.forename: Ronald
authors.initials: R
authors.lastname: Conaway
authors.forename: Jean Marc
authors.initials: JM
authors.lastname: Egly
mesh_terms: D004249:DNA Damage; D004265:DNA Helicases / Q000302:isolation & purification / Q000502:physiology; D004260:DNA Repair / Q000235:genetics*; D045643:DNA Repair Enzymes / Q000302:isolation & purification / Q000502:physiology; D000075065:Elongin; D006147:Guanine / Q000031:analogs & derivatives / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D018384:Oxidative Stress; D000075223:Poly-ADP-Ribose Binding Proteins; D012319:RNA Polymerase II / Q000037:antagonists & inhibitors*; D013941:Thymine / Q000031:analogs & derivatives / Q000494:pharmacology; D014157:Transcription Factors / Q000302:isolation & purification / Q000502:physiology; D019473:Transcription Factors, TFII / Q000302:isolation & purification / Q000502:physiology; D014158:Transcription, Genetic / Q000187:drug effects; D035602:Transcriptional Elongation Factors / Q000302:isolation & purification / Q000502:physiology*; D014498:Uracil / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D000075065:Elongin; D000075223:Poly-ADP-Ribose Binding Proteins; D014157:Transcription Factors; D019473:Transcription Factors, TFII; D035602:Transcriptional Elongation Factors; C018648:transcription factor S-II; C029389:thymine glycol; C045975:5-hydroxyuracil; C024829:8-hydroxyguanine; D014498:Uracil; D006147:Guanine; D012319:RNA Polymerase II; D004265:DNA Helicases; C078673:ERCC6 protein, human; C056604:transcription factor TFIIF; D045643:DNA Repair Enzymes; D013941:Thymine