A DNA-translocating Snf2 Molecular Motor: Saccharomyces cerevisiae Rdh54 Displays Processive Translocation and Extrudes DNA Loops

Tekkatte Krishnamurthy Prasad, Ragan B. Robertson, Mari Liis Visnapuu, Peter Chi, Patrick Sung, Eric C. Greene

Research output: Contribution to journalArticle

54 Scopus citations

Abstract

We have used total internal reflection fluorescence microscopy (TIRFM) to investigate the characteristics of the yeast homologous recombination factor Rdh54 on DNA. Our results demonstrate translocation of Rdh54 on DNA and extrusion of DNA loops by Rdh54 in an ATP hydrolysis-dependent manner. The translocating Rdh54 was highly processive and displayed a variety of behavior, including variations in translocation rate and distance, pauses, and reversals. We provide evidence that the DNA loops generated encompass an average of 6 kb, and Rdh54 often abruptly releases the extruded DNA. Rdh54 forms a multimeric complex, which we speculate has at least two functionally distinct DNA-binding sites, one of which enables translocation while the other remains anchored to another DNA locale. Our work, together with other recent studies, suggests that translocation-coupled DNA loop extrusion is a common mechanistic feature among the Snf2-family of chromatin-remodeling proteins.

Original languageEnglish (US)
Pages (from-to)940-953
Number of pages14
JournalJournal of Molecular Biology
Volume369
Issue number4
DOIs
StatePublished - Jun 15 2007
Externally publishedYes

Keywords

  • DNA curtain
  • homologous recombination
  • Rdh54
  • single-molecule
  • TIRFM

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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