selective incision of the - -methyl-formamidopyrimidine anomer by escherichia coli endonuclease iv选择性的切口,-methyl-formamidopyrimidine异头物由大肠杆菌核酸内切酶iv.pdfVIP

SAGE-Hindawi Access to Research Journal of Nucleic Acids Volume 2010, Article ID 850234, 10 pages doi:10.4061/2010/850234 Research Article Selective Incision of the α-N 5-Methyl-Formamidopyrimidine Anomer by Escherichia coli Endonuclease IV Plamen P. Christov, Surajit Banerjee, Michael P. Stone, and Carmelo J. Rizzo Departments of Chemistry and Biochemistry, Center in Molecular Toxicology, Vanderbilt University, VU Station B 351822, Nashville, N 37235-1822, USA Correspondence should be addressed to Carmelo J. Rizzo, c.j.rizzo@ Received 13 April 2010; Accepted 4 June 2010 Academic Editor: Ashis Basu Copyright © 2010 Plamen P. Christov et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Formamidopyrimidines (Fapy) lesions result from ring opening of the imidazole portion of purines. Fapy lesions can isomerize from the natural β -anomeric stereochemistry to the α-configuration. We have unambiguously demonstrated that the α-methyl- Fapy-dG (MeFapy-dG) lesion is a substrate for Escherichia coli Endonuclease IV (Endo IV). Treatment of a MeFapy-dG-containing 24 mer duplex with Endo IV resulted in 36–40% incision. The catalytic efficiency of the incision was comparable to that of α-dG in the same duplex sequence. The α- and β -MeFapy-dG anomers equilibrate to ∼21 : 79 ratio over ∼3 days. Related studies with a duplex containing the α-Fapy-dG lesion derived from aflatoxin B1 epoxide (α-AFB-Fapy-dG) showed only low levels of incision. It is hypothesized that the steric bulk of the aflatoxin moiety interferes with the binding of the substrate to Endo IV and the incision chemistry. 1. Introduction