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J. Biol. Chem., Vol. 276, Issue 48, 45367-45371, November 30, 2001
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From the 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.
Effect of Thymine Glycol on Transcription Elongation by T7 RNA
Polymerase and Mammalian RNA Polymerase II*
,,,¶
Department of Biological Sciences, Stanford
University, Stanford, California 94305-5020 and
§ Department of Biochemistry, Emory University School of
Medicine, Atlanta, Georgia 30322
*
This work was supported by Grant CA-77712 from the NCI,
National Institutes of Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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