Crystal and solution structures of d(CGC[e6G]AATTCGCG)-drug complexes reveal conformational polymorphism of O6-ethyl-guanine:cytosine base pair.
O6-ethyl-guanine (e6G) is a relatively persistent alkylation lesion caused by the exposure of DNA to carcinogen N-ethyl-N-nitrosourea. We have studied the structural consequences of the e6G incorporation in DNA by X-ray crystallography and NMR. We have obtained crystals of the modified DNA dodecamer d(CGC[e6G]AATTCGCG) complexed to several minor groove binding drugs including Hoechst 33258, Hoechst 33342, netropsin, and SN6999. The space group of the crystals from those complexes is P2(1)2(1)2(1). However the crystal structure of the SN6999 complex is not isomorphous to that from the other three complexes. In all four refined crystal structures the drugs bind in the narrow minor groove at or close to the central AATT region of the dodecamer B-DNA duplex. The DNA conformation is influenced by the binding of drugs. The eight independent e6G:C base pairs have a conformation ranging from one with three-centered hydrogen bonds between the bases to a wobble conformation with two hydrogen bonds. The ethyl group of the eight e6G bases is mostly in the proximal orientation to N7. Our 1D and 2D-NMR studies of the same (free) dodecamer reveal that the e6G:C base pairs in the duplex are likely to adopt a wobble conformation in solution. Those results suggest that the e6G:C base pair has a dynamic equilibrium among various conformations, which may present an ambiguous signal to cells. In contrast, the e6G:T base pair adopts a Watson-Crick-like conformation. This may be a plausible explanation of why thymine is found preferentially incorporated across the e6G during replication.
Sriram
M
M
Department of Cell and Structural Biology, University of Illinois at Urbana-Champaign 61801.
Yang
D
D
Gao
Y G
YG
Wang
A H
AH
D001482:Base Composition; D001483:Base Sequence; D003460:Crystallization; D003596:Cytosine / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D006147:Guanine / Q000031:analogs & derivatives* / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D004364:Pharmaceutical Preparations / Q000737:chemistry*
D004364:Pharmaceutical Preparations; C007673:6-ethylguanine; D006147:Guanine; D003596:Cytosine; D004247:DNA