5'-Single-stranded/duplex DNA junctions are loading sites for E. coli UvrD translocase

Eric J. Tomko, Haifeng Jia, Jeehae Park, Nasib K. Maluf, Taekjip Ha, Timothy M. Lohman

Research output: Contribution to journalArticle

Abstract

Escherichia coli UvrD is a 3'-5' superfamily 1A helicase/ translocase involved in a variety of DNA metabolic processes. UvrD can function either as a helicase or only as an singlestranded DNA (ssDNA) translocase. The switch between these activities is controlled in vitro by the UvrD oligomeric state; a monomer has ssDNA translocase activity, whereas at least a dimer is needed for helicase activity. Although a 30- ssDNA partial duplex provides a high-affinity site for a UvrD monomer, here we show that a monomer also binds with specificity to DNA junctions possessing a 50-ssDNA flanking region and can initiate translocation from this site. Thus, a 50-ss-duplex DNA junction can serve as a high-affinity loading site for the monomeric UvrD translocase, whereas a 30-ss-duplex DNA junction inhibits both translocase and helicase activity of the UvrD monomer. Furthermore, the 2B subdomain of UvrD is important for this junction speci- ficity. This highlights a separation of helicase and translocase function for UvrD and suggests that a monomeric UvrD translocase can be loaded at a 50-ssDNA junction when translocation activity alone is needed.

Original languageEnglish (US)
Pages (from-to)3826-3839
Number of pages14
JournalEMBO Journal
Volume29
Issue number22
DOIs
StatePublished - Nov 17 2010

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Keywords

  • DNA junctions
  • fluorescence
  • helicase
  • kinetics
  • translocase

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Tomko, E. J., Jia, H., Park, J., Maluf, N. K., Ha, T., & Lohman, T. M. (2010). 5'-Single-stranded/duplex DNA junctions are loading sites for E. coli UvrD translocase. EMBO Journal, 29(22), 3826-3839. https://doi.org/10.1038/emboj.2010.242