Effect of thymine glycol on transcription elongation by T7 RNA polymerase and mammalian RNA polymerase II.
Thymine glycols are formed in DNA by exposure to ionizing radiation or oxidative stress. Although these lesions are repaired by the base excision repair pathway, they have been shown also to be subject to transcription-coupled repair. A current model for transcription-coupled repair proposes that RNA polymerase II arrested at a DNA lesion provides a signal for recruitment of the repair enzymes to the lesion site. Here we report the effect of thymine glycol on transcription elongation by T7 RNA polymerase and RNA polymerase II from rat liver. DNA substrates containing a single thymine glycol located either in the transcribed or nontranscribed strand were used to carry out in vitro transcription. We found that thymine glycol in the transcribed strand blocked transcription elongation by T7 RNA polymerase approximately 50% of the time but did not block RNA polymerase II. Thymine glycol in the nontranscribed strand did not affect transcription by either polymerase. These results suggest that arrest of RNA polymerase elongation by thymine glycol is not necessary for transcription-coupled repair of this lesion. Additional factors that recognize and bind thymine glycol in DNA may be required to ensure RNA polymerase arrest and the initiation of transcription-coupled repair in vivo.
Tornaletti
S
S
Department of Biological Sciences, Stanford University, Stanford, California 94305-5020, USA.
Maeda
L S
LS
Lloyd
D R
DR
Reines
D
D
Hanawalt
P C
PC
D000818:Animals; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000378:metabolism; D004270:DNA, Circular / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D009841:Oligonucleotides / Q000494:pharmacology; D018384:Oxidative Stress; D010766:Phosphorylation; D012313:RNA / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism*; D051381:Rats; D013941:Thymine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D013997:Time Factors; D014158:Transcription, Genetic; D014764:Viral Proteins
D004270:DNA, Circular; D009841:Oligonucleotides; D014764:Viral Proteins; C029389:thymine glycol; D012313:RNA; D004247:DNA; D012319:RNA Polymerase II; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D013941:Thymine