DNA Polymerase-Parental DNA Interaction Is Essential for Helicase-Polymerase Coupling during Bacteriophage T7 DNA Replication

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dc.citation.articleNumber1342en_US
dc.citation.issueNumber3en_US
dc.citation.journalTitleInternational Journal of Molecular Sciencesen_US
dc.citation.volumeNumber23en_US
dc.contributor.authorLo, Chen-Yuen_US
dc.contributor.authorGao, Yangen_US
dc.date.accessioned2022-02-04T19:19:12Zen_US
dc.date.available2022-02-04T19:19:12Zen_US
dc.date.issued2022en_US
dc.description.abstractDNA helicase and polymerase work cooperatively at the replication fork to perform leading-strand DNA synthesis. It was believed that the helicase migrates to the forefront of the replication fork where it unwinds the duplex to provide templates for DNA polymerases. However, the molecular basis of the helicase-polymerase coupling is not fully understood. The recently elucidated T7 replisome structure suggests that the helicase and polymerase sandwich parental DNA and each enzyme pulls a daughter strand in opposite directions. Interestingly, the T7 polymerase, but not the helicase, carries the parental DNA with a positively charged cleft and stacks at the fork opening using a β-hairpin loop. Here, we created and characterized T7 polymerases each with a perturbed β-hairpin loop and positively charged cleft. Mutations on both structural elements significantly reduced the strand-displacement synthesis by T7 polymerase but had only a minor effect on DNA synthesis performed against a linear DNA substrate. Moreover, the aforementioned mutations eliminated synergistic helicase-polymerase binding and unwinding at the DNA fork and processive fork progressions. Thus, our data suggested that T7 polymerase plays a dominant role in helicase-polymerase coupling and replisome progression.en_US
dc.identifier.citationLo, Chen-Yu and Gao, Yang. "DNA Polymerase-Parental DNA Interaction Is Essential for Helicase-Polymerase Coupling during Bacteriophage T7 DNA Replication." <i>International Journal of Molecular Sciences,</i> 23, no. 3 (2022) MDPI: https://doi.org/10.3390/ijms23031342.en_US
dc.identifier.digitalijms-23-01342en_US
dc.identifier.doihttps://doi.org/10.3390/ijms23031342en_US
dc.identifier.urihttps://hdl.handle.net/1911/111972en_US
dc.language.isoengen_US
dc.publisherMDPIen_US
dc.rightsThis is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly citeden_US
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en_US
dc.titleDNA Polymerase-Parental DNA Interaction Is Essential for Helicase-Polymerase Coupling during Bacteriophage T7 DNA Replicationen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
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