Effect of thymine glycol on transcription elongation by T7 RNA polymerase and mammalian RNA polymerase II

• Authors: Silvia Tornaletti, Stanford University; Lauren S. Maeda, Stanford University; Daniel R. Lloyd, Stanford University; Daniel Reines, Emory University; Philip C. Hanawalt, Stanford University
• Journal Title: Journal of Biological Chemistry
• Volume: Volume 276, Number 48
• Publisher: American Society for Biochemistry and Molecular Biology | 2001-11-30, Pages 45367-45371
• Type of Work: Article | Final Publisher PDF
• Publisher DOI: http://doi.org/10.1074/jbc.M105282200
• Subjects: Chemistry, Biochemistry; Biology, Molecular
• Keywords: Science & Technology; Life Sciences & Biomedicine; Biochemistry & Molecular Biology; CYCLOBUTANE PYRIMIDINE DIMER; DNA-BASE DAMAGES; COUPLED REPAIR; ESCHERICHIA-COLI; COCKAYNE-SYNDROME; UREA RESIDUES; HUMAN-CELLS; DUPLEX DNA; STRAND; GENE

Abstract

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 ∼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.