title: Intrinsic stability and oligomerization dynamics of DNA processivity clamps.
abstract: Sliding clamps are ring-shaped oligomeric proteins that are essential for processive deoxyribonucleic acid replication. Although crystallographic structures of several clamps have been determined, much less is known about clamp structure and dynamics in solution. Here, we characterized the intrinsic solution stability and oligomerization dynamics of the homodimeric Escherichia coli β and the homotrimeric Saccharomyces cerevisiae proliferating cell nuclear antigen (PCNA) clamps using single-molecule approaches. We show that E. coli β is stable in solution as a closed ring at concentrations three orders of magnitude lower than PCNA. The trimeric structure of PCNA results in slow subunit association rates and is largely responsible for the lower solution stability. Despite this large difference, the intrinsic lifetimes of the rings differ by only one order of magnitude. Our results show that the longer lifetime of the E. coli β dimer is due to more prominent electrostatic interactions that stabilize the subunit interfaces.
authors.affiliation: Department of Chemistry and Biochemistry and Biodesign Institute, Arizona State University, Tempe, AZ 85287-5601, USA.
authors.forename: Jennifer K
authors.initials: JK
authors.lastname: Binder
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32610-0245, USA.
authors.forename: Lauren G
authors.initials: LG
authors.lastname: Douma
authors.affiliation: Department of Chemistry and Biochemistry and Biodesign Institute, Arizona State University, Tempe, AZ 85287-5601, USA.
authors.forename: Suman
authors.initials: S
authors.lastname: Ranjit
authors.affiliation: Department of Chemistry and Biochemistry and Biodesign Institute, Arizona State University, Tempe, AZ 85287-5601, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Kanno
authors.affiliation: Department of Chemistry and Biochemistry and Biodesign Institute, Arizona State University, Tempe, AZ 85287-5601, USA.
authors.forename: Manas
authors.initials: M
authors.lastname: Chakraborty
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32610-0245, USA lbloom@ufl.edu.
authors.forename: Linda B
authors.initials: LB
authors.lastname: Bloom
authors.affiliation: Department of Chemistry and Biochemistry and Biodesign Institute, Arizona State University, Tempe, AZ 85287-5601, USA mlevitus@asu.edu.
authors.forename: Marcia
authors.initials: M
authors.lastname: Levitus
mesh_terms: D004258:DNA Polymerase III / Q000737:chemistry / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry / Q000378:metabolism*; D055503:Protein Multimerization; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D029968:Escherichia coli Proteins; D018809:Proliferating Cell Nuclear Antigen; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; C101139:beta subunit, DNA polymerase III; D004258:DNA Polymerase III
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