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O sup 6 -ethylguanine carcinogenic lesions in DNA: An NMR study of O sup 6 etGter dot T pairing in dodecanucloetide duplexes

Journal Article · · Biochemistry; (USA)
DOI:https://doi.org/10.1021/bi00441a008· OSTI ID:5432900

High-resolution two-dimensional NMR studies are reported on the self-complementary d-(C1-G2-C3-{und O{sup 6}etG4}-A5-G6-C7-T8-{und T9}-G10-C11-G12) duplex (designated O{sup 6}etG{center dot}T 12-mer) containing two symmetrically related O{sup 6}etG{center dot}T lesion sites located four base pairs in from either end of the duplex. Parallel studies were undertaken on a related sequence containing O{sup 6}meG{center dot}T lesion sites (designated O{sup 6}meG{center dot}T 12-mer) in order to evaluate the influence of the size of the alkyl substituent on the structure of the duplex and were undertaken on a related sequence containing G{center dot}T mismatch sites (designated G{center dot}T 12-mer duplex), which served as the control duplex. The exchangeable and nonexchangeable proton and the phosphorus nuclei have been assigned from an analysis of two-dimensional nuclear Overhauser enhancement (NOE) and correlated spectra of the O{sup 6}etG{center dot}T 12-mer, O{sup 6}meG{center dot}T 12-mer, and G{center dot}T 12-mer duplexes in H{sub 2}O and D{sub 2}O solutions. The distance connectivities observed in the NOESY spectra of the O{sup 6}alkG{center dot}T 12-mer duplexes establish that the helix is right-handed and all of the bases adopt an anti conformation of the glycosidic torsion angle including the O{sup 6}alkG4 and T9 bases at the lesion site. These observations establish that the O{sup 6}alkG4 and T9 residues are stacked into the duplex and that the O{sup 6}CH{sub 3} and O{sup 6}CH{sub 2}CH{sub 3} groups of O{sup 6}alkG4 adopt a syn orientation with respect to the N{sup 1} of the alkylated guanine. Since the O{sup 6}-alkyl group adopts a syn orientation, the separation between the O{sup 6} of O{sup 6}alkG4 and the O{sup 4} of T9 in the major groove is increased, preventing the formation of a short hydrogen bond between the N{sup 1} ring nitrogen of O{sup 6}alkG4 and the imino proton of T9.

OSTI ID:
5432900
Journal Information:
Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 28:15; ISSN 0006-2960; ISSN BICHA
Country of Publication:
United States
Language:
English