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dc.contributor.authorKhaki, Ali R.
dc.contributor.authorField, Cassandra
dc.contributor.authorMalik, Shuja Shafi
dc.contributor.authorNiedziela-Majka, Anita
dc.contributor.authorLeavitt, Stephanie A.
dc.contributor.authorWang, Ruth
dc.contributor.authorHung, Magdeleine
dc.contributor.authorSakowicz, Roman
dc.contributor.authorBrendza, Katherine M.
dc.contributor.authorFischer, Christopher J.
dc.date.accessioned2017-03-27T20:57:46Z
dc.date.available2017-03-27T20:57:46Z
dc.date.issued2010-05-06
dc.identifier.citationKhaki, Ali R., Cassandra Field, Shuja Malik, Anita Niedziela-Majka, Stephanie A. Leavitt, Ruth Wang, Magdeleine Hung, Roman Sakowicz, Katherine M. Brendza, and Christopher J. Fischer. "The Macroscopic Rate of Nucleic Acid Translocation by Hepatitis C Virus Helicase NS3h Is Dependent on Both Sugar and Base Moieties." Journal of Molecular Biology 400.3 (2010): 354-78.en_US
dc.identifier.urihttp://hdl.handle.net/1808/23490
dc.description.abstractThe NS3 helicase (NS3h) of hepatitis C virus (HCV) is a 3′ to 5′ SF2 RNA and DNA helicase that is essential for the replication of HCV. We have examined the kinetic mechanism of translocation of NS3h along single-stranded nucleic acid with bases rU, dU and dT and have found that the macroscopic rate of translocation is dependent upon both the base and sugar moieties of the nucleic acid, with approximate macroscopic translocation rates of 3 nt/s (oligo-dT), 35 nt/s (oligo-dU), and 42 nt/s (oligo-rU), respectively. We found a strong correlation between the macroscopic translocation rates and the binding affinity of the translocating NS3h protein to the respective substrates such that weaker affinity corresponded to faster translocation. The values of K0.5 for NS3h translocation at a saturating ATP concentration are: (3.3 ± 0.4) μM nucleotide (poly-dT), (27 ± 2) μM nucleotide (poly-dU), and (36 ± 2) μM nucleotide (poly-rU). Furthermore, the results of isothermal titration of NS3h with these oligonucleotides suggest that differences in TΔS° are the principal source of the differences in the affinity of NS3h binding to these substrates. Interestingly, despite the differences in macroscopic translocation rates and binding affinities, the ATP coupling stoichiometry for NS3h translocation was identical for all three substrates, ~0.5 ATP molecules consumed per nucleotide translocated. This similar periodicity of ATP consumption implies a similar mechanism for NS3h translocation along RNA and DNA substrates.en_US
dc.publisherElsevieren_US
dc.rightsThis is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License 3.0 (CC BY-NC-ND 3.0 US), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.en_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/
dc.subjectNS3 helicaseen_US
dc.subjectTranslocationen_US
dc.subjectNucleic aciden_US
dc.subjectATPaseen_US
dc.subjectMechanismen_US
dc.titleThe Macroscopic Rate of Nucleic Acid Translocation by Hepatitis C virus Helicase NS3h is Dependent on Both the Sugar and Base Moietiesen_US
dc.typeArticleen_US
kusw.kuauthorFischer, Christopher J.
kusw.kudepartmentPhysics and Astronomyen_US
dc.identifier.doi10.1016/j.jmb.2010.04.065en_US
kusw.oaversionScholarly/refereed, author accepted manuscripten_US
kusw.oapolicyThis item meets KU Open Access policy criteria.en_US
dc.rights.accessrightsopenAccess


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This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License 3.0 (CC BY-NC-ND 3.0 US), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
Except where otherwise noted, this item's license is described as: This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License 3.0 (CC BY-NC-ND 3.0 US), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.