Base excision repair in a glucocorticoid response element: effect of glucocorticoid receptor binding.

The Journal of Biological Chemistry
Angela K HinzMichael Smerdon

Abstract

DNA repair takes place in the context of chromatin. Previous studies showed that histones impair base excision repair (BER) of modified bases at both the excision and synthesis steps. We examined BER of uracil in a glucocorticoid response element (GRE) complexed with the glucocorticoid receptor DNA binding domain (GR-DBD). Five substrates were designed, each containing a unique C-->U substitution within the mouse mammary tumor virus promoter, one located within each GRE half-site and the others located outside the GRE. To examine distinct steps of BER, DNA cleavage by uracil-DNA glycosylase and Ape1 endonuclease was used to assess initiation, dCTP incorporation by DNA polymerase (pol) beta was used to measure repair synthesis, and DNA ligase I was used to seal the nick. For uracil sites within the GRE, there was a reduced rate of uracil-DNA glycosylase/Ape1 activity following GR-DBD binding. Cleavage in the right half-site, with higher GR-DBD binding affinity, was reduced approximately 5-fold, whereas cleavage in the left half-site was reduced approximately 3.8-fold. Conversely, uracil-directed cleavage outside the GRE was unaffected by GR-DBD binding. Surprisingly, there was no reduction in the rate of pol beta synthesis or DN...Continue Reading

References

Mar 1, 1992·The Journal of Steroid Biochemistry and Molecular Biology·K Dahlman-WrightJ A Gustafsson
Jan 1, 1996·Proteins·T C Bishop, K Schulten
Jan 10, 1997·The Journal of Biological Chemistry·P R StraussS H Wilson
Jun 17, 1997·Biochemistry·I D NichollM K Kenny
Aug 1, 1998·The Journal of Biological Chemistry·E K DimitriadisS H Wilson
Aug 8, 1998·The Journal of Biological Chemistry·D K SrivastavaS H Wilson
Aug 24, 1999·Nucleic Acids Research·R Ghosh, D L Mitchell
Mar 4, 2000·Nature Structural Biology·S H Wilson, T A Kunkel
Aug 1, 2000·Mutation Research·A Memisoglu, L Samson
Oct 4, 2000·Nature Structural Biology·K S WilsonY Nakamura
Nov 2, 2002·The EMBO Journal·Hilde NilsenAlain Verreault
Jun 12, 2003·Proceedings of the National Academy of Sciences of the United States of America·Brian C BeardMichael J Smerdon
May 26, 2005·Mutation Research·Youngho Kwon, Michael J Smerdon
Jun 9, 2005·Mutation Research·Joy L HuffmanJohn A Tainer
Nov 25, 2006·DNA Repair·Grigory L Dianov, Jason L Parsons
Dec 18, 2007·Molecular Cell·Philip C Hanawalt

❮ Previous
Next ❯

Methods Mentioned

BETA
glycosylase
electrophoresis
Assay

Software Mentioned

ImageQuant
PyMOL

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.