Intrinsic stability and oligomerization dynamics of DNA processivity clamps

Jennifer K. Binder, Lauren G. Douma, Suman Ranjit, David M. Kanno, Manas Chakraborty, Linda B. Bloom, Marcia Levitus

Research output: Contribution to journalArticle

11 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)6476-6486
Number of pages11
JournalNucleic acids research
Volume42
Issue number10
DOIs
StatePublished - 2014

ASJC Scopus subject areas

  • Genetics

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    Binder, J. K., Douma, L. G., Ranjit, S., Kanno, D. M., Chakraborty, M., Bloom, L. B., & Levitus, M. (2014). Intrinsic stability and oligomerization dynamics of DNA processivity clamps. Nucleic acids research, 42(10), 6476-6486. https://doi.org/10.1093/nar/gku255