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dc.contributor.authorWilliamson, Adele Kim
dc.contributor.authorGrgic, Miriam
dc.contributor.authorLeiros, Hanna-Kirsti S.
dc.date.accessioned2019-03-19T18:19:11Z
dc.date.available2019-03-19T18:19:11Z
dc.date.issued2018-07-11
dc.description.abstractDNA ligases join breaks in the phosphodiester backbone of DNA by catalysing the formation of bonds between opposing 5′P and 3′OH ends in an adenylation-dependent manner. Catalysis is accompanied by reorientation of two core domains to provide access to the active site for cofactor utilization and enable substrate binding and product release. The general paradigm is that DNA ligases engage their DNA substrate through complete encirclement of the duplex, completed by inter-domain kissing contacts via loops or additional domains. The recent structure of a minimal Lig E-type DNA ligase, however, implies it must use a different mechanism, as it lacks any domains or loops appending the catalytic core which could complete encirclement. In the present study, we have used a structure-guided mutagenesis approach to investigate the role of conserved regions in the Lig E proteins with respect to DNA binding. We report the structure of a Lig-E type DNA ligase bound to the nicked DNA-adenylate reaction intermediate, confirming that complete encirclement is unnecessary for substrate engagement. Biochemical and biophysical measurements of point mutants to residues implicated in binding highlight the importance of basic residues in the OB domain, and inter-domain contacts to the linker.en_US
dc.description.sponsorshipThe publication fund at the University of Tromsøen_US
dc.descriptionSource at <a href=https://doi.org/10.1093/nar/gky622> https://doi.org/10.1093/nar/gky622</a>.en_US
dc.identifier.citationWilliamson, A.K., Grgic, M. & Leiros H.-K.S. (2018). DNA binding with a minimal scaffold: structure-function analysis of Lig E DNA ligases. <i>Nucleic Acids Research, 46</i>(16), 8616-8629. https://doi.org/10.1093/nar/gky622en_US
dc.identifier.cristinIDFRIDAID 1623616
dc.identifier.doi10.1093/nar/gky622
dc.identifier.issn0305-1048
dc.identifier.issn1362-4962
dc.identifier.urihttps://hdl.handle.net/10037/15029
dc.language.isoengen_US
dc.publisherOxford University Pressen_US
dc.relation.journalNucleic Acids Research
dc.relation.projectIDinfo:eu-repo/grantAgreement/RCN/BIOTEK2021/244247/Norway/Engineering efficient DNA ligases for improved Next-Generation-Sequencing/BCSS/en_US
dc.relation.projectIDinfo:eu-repo/grantAgreement/RCN/SYNKNØYT/247732/Norway/Reisestøtte, synkrotron- og nøytronforskning, 2015-2017//en_US
dc.rights.accessRightsopenAccessen_US
dc.subjectVDP::Mathematics and natural science: 400::Chemistry: 440en_US
dc.subjectVDP::Matematikk og Naturvitenskap: 400::Kjemi: 440en_US
dc.titleDNA binding with a minimal scaffold: structure-function analysis of Lig E DNA ligasesen_US
dc.typeJournal articleen_US
dc.typeTidsskriftartikkelen_US
dc.typePeer revieweden_US


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