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dc.contributor.authorLee, Seung-Joo
dc.contributor.authorZhu, Bin
dc.contributor.authorHamdan, Samir
dc.contributor.authorRichardson, Charles C.
dc.date.accessioned2014-08-27T09:51:49Z
dc.date.available2014-08-27T09:51:49Z
dc.date.issued2010-03-28
dc.identifier.citationLee S-J, Zhu B, Hamdan SM, Richardson CC (2010) Mechanism of sequence-specific template binding by the DNA primase of bacteriophage T7. Nucleic Acids Research 38: 4372-4383. doi:10.1093/nar/gkq205.
dc.identifier.issn03051048
dc.identifier.pmid20350931
dc.identifier.doi10.1093/nar/gkq205
dc.identifier.urihttp://hdl.handle.net/10754/325449
dc.description.abstractDNA primases catalyze the synthesis of the oligoribonucleotides required for the initiation of lagging strand DNA synthesis. Biochemical studies have elucidated the mechanism for the sequence-specific synthesis of primers. However, the physical interactions of the primase with the DNA template to explain the basis of specificity have not been demonstrated. Using a combination of surface plasmon resonance and biochemical assays, we show that T7 DNA primase has only a slightly higher affinity for DNA containing the primase recognition sequence (5'-TGGTC-3') than for DNA lacking the recognition site. However, this binding is drastically enhanced by the presence of the cognate Nucleoside triphosphates (NTPs), Adenosine triphosphate (ATP) and Cytosine triphosphate (CTP) that are incorporated into the primer, pppACCA. Formation of the dimer, pppAC, the initial step of sequence-specific primer synthesis, is not sufficient for the stable binding. Preformed primers exhibit significantly less selective binding than that observed with ATP and CTP. Alterations in subdomains of the primase result in loss of selective DNA binding. We present a model in which conformational changes induced during primer synthesis facilitate contact between the zinc-binding domain and the polymerase domain. The Author(s) 2010. Published by Oxford University Press.
dc.language.isoen
dc.publisherOxford University Press (OUP)
dc.rightsThis is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
dc.rights.urihttp://creativecommons.org/licenses/by-nc/2.5
dc.subjectadenosine triphosphate
dc.subjectcytidine triphosphate
dc.subjectDNA primase
dc.subjectnucleoside triphosphatase
dc.subjectbacteriophage T7
dc.subjectbinding affinity
dc.subjectbinding site
dc.subjectcontrolled study
dc.subjectDNA binding
dc.subjectDNA determination
dc.subjectDNA sequence
dc.subjectDNA synthesis
dc.subjectDNA template
dc.subjectgene function
dc.subjectsurface plasmon resonance
dc.subjectAdenosine Triphosphate
dc.subjectBacteriophage T7
dc.subjectBase Sequence
dc.subjectBinding Sites
dc.subjectCytidine Triphosphate
dc.subjectDNA Primase
dc.subjectDNA, Single-Stranded
dc.subjectDNA-Binding Proteins
dc.subjectModels, Molecular
dc.subjectOligoribonucleotides
dc.subjectProtein Binding
dc.subjectProtein Structure, Tertiary
dc.subjectTemplates, Genetic
dc.titleMechanism of sequence-specific template binding by the DNA primase of bacteriophage T7
dc.typeArticle
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Division
dc.identifier.journalNucleic Acids Research
dc.identifier.pmcidPMC2910064
dc.eprint.versionPublisher's Version/PDF
dc.contributor.institutionDepartment of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, United States
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)
kaust.personHamdan, Samir
refterms.dateFOA2018-06-13T16:06:48Z


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This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Except where otherwise noted, this item's license is described as This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.