The mutation frequency of 8-oxo-7,8-dihydroguanine (8-oxodG) situated in a multiply damaged site: comparison of a single and two closely opposed 8-oxodG in Escherichia coli

DNA Repair
Svitlana MalyarchukL Harrison

Abstract

A multiply damaged site (MDS) is defined as > or =2 lesions within a distance of 10-15 base pairs (bp). MDS generated by ionizing radiation contain oxidative base damage, and in vitro studies have indicated that if the base damage is <3bp apart, repair of one lesion is inhibited until repair of the lesion in the opposite strand is completed. Inhibition of repair could result in an increase in the mutation frequency of the base damage. We have designed an assay to determine whether a closely opposed lesion causes an increase in adenine insertion opposite an 8-oxodG in bacteria. We have positioned the MDS (an 8-oxodG in the transcribed strand and a second 8-oxodG immediately 5' to this lesion in the non-transcribed strand) within the firefly luciferase coding region. During two rounds of replication, insertion of adenine opposite the 8-oxodG in the transcribed (T) or non-transcribed (NT) strand results in a translation termination codon at position 444 or 445, respectively. The truncated luciferase protein is inactive. We have generated double-stranded oligonucleotides that contain no damage, each single 8-oxodG or the MDS. Each double-stranded molecule was ligated into the reporter vector and the ligation products transformed in...Continue Reading

References

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Jan 5, 2000·Proceedings of the National Academy of Sciences of the United States of America·B M SutherlandJ Laval
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Jun 1, 2002·Pathophysiology : the Official Journal of the International Society for Pathophysiology·Lynn Harrison, Svitlana Malyarchuk
Aug 2, 2002·Molecular Cell·Youri I PavlovThomas A Kunkel

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Citations

Jul 31, 2003·Nucleic Acids Research·Dwain I D'souza, Lynn Harrison
Jan 14, 2005·Nucleic Acids Research·Grégory Eot-HoullierEvelyne Sage
May 1, 2007·Nucleic Acids Research·Grégory Eot-HoullierEvelyne Sage
May 27, 2009·Nucleic Acids Research·Sophie BellonPeter O'Neill
Feb 12, 2009·International Journal of Radiation Biology·Peter O'Neill, Peter Wardman
Apr 5, 2005·Journal of Computational Chemistry·Hirofumi FujimotoHideaki Maekawa
Dec 28, 2010·Mutation Research·Evelyne Sage, Lynn Harrison
Aug 5, 2010·Radiation Research·Jennifer A AndersonPeter O'Neill
May 21, 2013·Radiation Research·Alexandros G GeorgakilasRobert D Stewart
Feb 23, 2012·Cancer Letters·Karin Magnander, Kecke Elmroth
Dec 7, 2010·Mutation Research·Laura J EcclesMartine E Lomax
Dec 9, 2008·International Journal of Radiation Biology·Paban K AgrawalaAnnie Chetioui
Feb 17, 2009·Journal of Radiation Research·Naoya ShikazonoAkinari Yokoya
Nov 10, 2013·Cancers·Amanda F SwindallEddy S Yang
Oct 6, 2009·DNA Repair·Svitlana MalyarchukLynn Harrison

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