Dynamics of the ssDNA Recognition by the RepA Hexameric Helicase of Plasmid RSF1010: Analyses Using Fluorescence Stopped-Flow Intensity and Anisotropy Methods |
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| Authors: | Iraida E. Andreeva Wlodzimierz Bujalowski |
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| Affiliation: | Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA Department of Obstetrics and Gynecology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA Sealy Center for Structural Biology and Sealy Center for Cancer Cell Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA |
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| Abstract: | The kinetic mechanism of the single-stranded DNA (ssDNA) recognition by the RepA hexameric replicative helicase of the plasmid RSF1010 and the nature of formed intermediates, in the presence of the ATP nonhydrolyzable analog, β,γ-imidoadenosine-5′-triphosphate (AMP-PNP), have been examined, using the fluorescence intensity and anisotropy stopped-flow and analytical ultracentrifugation methods. Association of the RepA hexamer with the ssDNA oligomers that engage the total DNA-binding site and exclusively the strong DNA-binding subsite is a minimum four-step mechanism |
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| Keywords: | AMP-PNP, β,γ-imidoadenosine-5&prime -triphosphate ssDNA, single-stranded DNA |
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