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

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dc.citation.articleNumber1342
dc.citation.issueNumber3
dc.citation.journalTitleInternational Journal of Molecular Sciences
dc.citation.volumeNumber23
dc.contributor.authorLo, Chen-Yu
dc.contributor.authorGao, Yang
dc.date.accessioned2022-02-04T19:19:12Z
dc.date.available2022-02-04T19:19:12Z
dc.date.issued2022
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.
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.
dc.identifier.digitalijms-23-01342
dc.identifier.doihttps://doi.org/10.3390/ijms23031342
dc.identifier.urihttps://hdl.handle.net/1911/111972
dc.language.isoeng
dc.publisherMDPI
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 cited
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.titleDNA Polymerase-Parental DNA Interaction Is Essential for Helicase-Polymerase Coupling during Bacteriophage T7 DNA Replication
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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