Download MendeleyKinetics, Structure, and Mechanism of 8-Oxo-7,8-dihydro-2'-deoxyguanosine Bypass by Human DNA Polymerase eta
Patra, A., Nagy, L.D., Zhang, Q., Su, Y., Muller, L., Guengerich, F.P., Egli, M.(2014) J Biol Chem 289: 16867-16882
- PubMed: 24759104
- DOI: https://doi.org/10.1074/jbc.M114.551820
- Primary Citation of Related Structures:
4O3N, 4O3O, 4O3P, 4O3Q, 4O3R, 4O3S - PubMed Abstract:
DNA damage incurred by a multitude of endogenous and exogenous factors constitutes an inevitable challenge for the replication machinery. Cells rely on various mechanisms to either remove lesions or bypass them in a more or less error-prone fashion. The latter pathway involves the Y-family polymerases that catalyze trans-lesion synthesis across sites of damaged DNA. 7,8-Dihydro-8-oxo-2'-deoxyguanosine (8-oxoG) is a major lesion that is a consequence of oxidative stress and is associated with cancer, aging, hepatitis, and infertility. We have used steady-state and transient-state kinetics in conjunction with mass spectrometry to analyze in vitro bypass of 8-oxoG by human DNA polymerase ¦Ç (hpol ¦Ç). Unlike the high fidelity polymerases that show preferential insertion of A opposite 8-oxoG, hpol ¦Ç is capable of bypassing 8-oxoG in a mostly error-free fashion, thus preventing GC¡úAT transversion mutations. Crystal structures of ternary hpol ¦Ç-DNA complexes and incoming dCTP, dATP, or dGTP opposite 8-oxoG reveal that an arginine from the finger domain assumes a key role in avoiding formation of the nascent 8-oxoG:A pair. That hpol ¦Ç discriminates against dATP exclusively at the insertion stage is confirmed by structures of ternary complexes that allow visualization of the extension step. These structures with G:dCTP following either 8-oxoG:C or 8-oxoG:A pairs exhibit virtually identical active site conformations. Our combined data provide a detailed understanding of hpol ¦Ç bypass of the most common oxidative DNA lesion.
Organizational Affiliation:
From the Department of Biochemistry and Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232 and.
