Primer terminal ribonucleotide alters the active site dynamics of DNA polymerase η and reduces DNA synthesis fidelity

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dc.citation.articleNumber102938en_US
dc.citation.issueNumber3en_US
dc.citation.journalTitleJournal of Biological Chemistryen_US
dc.citation.volumeNumber299en_US
dc.contributor.authorChang, Caleben_US
dc.contributor.authorLee Luo, Christieen_US
dc.contributor.authorEleraky, Sarahen_US
dc.contributor.authorLin, Aaronen_US
dc.contributor.authorZhou, Graceen_US
dc.contributor.authorGao, Yangen_US
dc.date.accessioned2023-03-10T19:04:10Zen_US
dc.date.available2023-03-10T19:04:10Zen_US
dc.date.issued2023en_US
dc.description.abstractDNA polymerases catalyze DNA synthesis with high efficiency, which is essential for all life. Extensive kinetic and structural efforts have been executed in exploring mechanisms of DNA polymerases, surrounding their kinetic pathway, catalytic mechanisms, and factors that dictate polymerase fidelity. Recent time-resolved crystallography studies on DNA polymerase η (Pol η) and β have revealed essential transient events during the DNA synthesis reaction, such as mechanisms of primer deprotonation, separated roles of the three metal ions, and conformational changes that disfavor incorporation of the incorrect substrate. DNA-embedded ribonucleotides (rNs) are the most common lesion on DNA and a major threat to genome integrity. While kinetics of rN incorporation has been explored and structural studies have revealed that DNA polymerases have a steric gate that destabilizes ribonucleotide triphosphate binding, the mechanism of extension upon rN addition remains poorly characterized. Using steady-state kinetics, static and time-resolved X-ray crystallography with Pol η as a model system, we showed that the extra hydroxyl group on the primer terminus does alter the dynamics of the polymerase active site as well as the catalysis and fidelity of DNA synthesis. During rN extension, Pol η error incorporation efficiency increases significantly across different sequence contexts. Finally, our systematic structural studies suggest that the rN at the primer end improves primer alignment and reduces barriers in C2′-endo to C3′-endo sugar conformational change. Overall, our work provides further mechanistic insights into the effects of rN incorporation on DNA synthesis.en_US
dc.identifier.citationChang, Caleb, Lee Luo, Christie, Eleraky, Sarah, et al.. "Primer terminal ribonucleotide alters the active site dynamics of DNA polymerase η and reduces DNA synthesis fidelity." <i>Journal of Biological Chemistry,</i> 299, no. 3 (2023) Elsevier: https://doi.org/10.1016/j.jbc.2023.102938.en_US
dc.identifier.digitalPIIS0021925823000704en_US
dc.identifier.doihttps://doi.org/10.1016/j.jbc.2023.102938en_US
dc.identifier.urihttps://hdl.handle.net/1911/114494en_US
dc.language.isoengen_US
dc.publisherElsevieren_US
dc.rightsThis is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).en_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/).en_US
dc.titlePrimer terminal ribonucleotide alters the active site dynamics of DNA polymerase η and reduces DNA synthesis fidelityen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
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