A pre-catalytic non-covalent step governs DNA polymerase β fidelity

Khadijeh S. Alnajjar, Ivan S. Krylov, Amirsoheil Negahbani, Pouya Haratipour, Boris A. Kashemirov, Ji Huang, Mariam Mahmoud, Charles E. McKenna, Myron F. Goodman, Joann B. Sweasy

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

DNA polymerase β (pol β) selects the correct deoxyribonucleoside triphosphate for incorporation into the DNA polymer. Mistakes made by pol β lead to mutations, some of which occur within specific sequence contexts to generate mutation hotspots. The adenomatous polyposis coli (APC) gene is mutated within specific sequence contexts in colorectal carcinomas but the underlying mechanism is not fully understood. In previous work, we demonstrated that a somatic colon cancer variant of pol β, K289M, misincorporates deoxynucleotides at significantly increased frequencies over wild-type pol β within a mutation hotspot that is present several times within the APC gene. Kinetic studies provide evidence that the rate-determining step of pol β catalysis is phosphodiester bond formation and suggest that substrate selection is governed at this step. Remarkably, we show that, unlike WT, a pre-catalytic step in the K289M pol β kinetic pathway becomes slower than phosphodiester bond formation with the APC DNA sequence but not with a different DNA substrate. Based on our studies, we propose that pre-catalytic conformational changes are of critical importance for DNA polymerase fidelity within specific DNA sequence contexts.

Original languageEnglish (US)
Pages (from-to)11839-11849
Number of pages11
JournalNucleic acids research
Volume47
Issue number22
DOIs
StatePublished - Dec 16 2019

ASJC Scopus subject areas

  • Genetics

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