Abstract
O6-alkylguanine-DNA alkyltransferase (AGT) is a single-cycle DNA repair enzyme that removes pro-mutagenic O6-alkylguanine adducts from DNA. Its functions with short single-stranded and duplex substrates have been characterized, but its ability to act on other DNA structures remains poorly understood. Here, we examine the functions of this enzyme on O6-methylguanine (6mG) adducts in the four-stranded structure of the human telomeric G-quadruplex. On a folded 22-nt G-quadruplex substrate, binding saturated at 2 AGT:DNA, significantly less than the ~ 5 AGT:DNA found with linear single-stranded DNAs of similar length, and less than the value found with the telomere sequence under conditions that inhibit quadruplex formation (4 AGT:DNA). Despite these differences, AGT repaired 6mG adducts located within folded G-quadruplexes, at rates that were comparable to those found for a duplex DNA substrate under analogous conditions. Repair was kinetically biphasic with the amplitudes of rapid and slow phases dependent on the position of the adduct within the G-quadruplex: in general, adducts located in the top or bottom tetrads of a quadruplex stack exhibited more rapid-phase repair than did adducts located in the inner tetrad. This distinction may reflect differences in the conformational dynamics of 6mG residues in G-quadruplex DNAs.
Document Type
Article
Publication Date
9-2-2014
Digital Object Identifier (DOI)
http://dx.doi.org/10.1093/nar/gku659
Funding Information
National Institutes of Health (NIH) [GM-070662 to M.G.F.]. Funding for open access charge: NIH [GM-070662 to M.G.F.].
Repository Citation
Hellman, Lance M.; Spear, Tyler J.; Koontz, Colton J.; Melikishvili, Manana; and Fried, Michael G., "Repair of O6-Methylguanine Adducts in Human Telomeric G-Quadruplex DNA by O6-Alkylguanine-DNA Alkyltransferase" (2014). Molecular and Cellular Biochemistry Faculty Publications. 75.
https://uknowledge.uky.edu/biochem_facpub/75
Notes/Citation Information
Published in Nucleic Acids Research, v. 42, no. 15, p. 9781-9791.
© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.