Saccharomyces cerevisiae Hrq1 requires a long 3'-tailed DNA substrate for helicase activity

Biochem Biophys Res Commun. 2012 Oct 26;427(3):623-8. doi: 10.1016/j.bbrc.2012.09.109. Epub 2012 Sep 28.

Abstract

RecQ helicases are well conserved proteins from bacteria to human and function in various DNA metabolism for maintenance of genome stability. Five RecQ helicases are found in humans, whereas only one RecQ helicase has been described in lower eukaryotes. However, recent studies predicted the presence of a second RecQ helicase, Hrq1, in fungal genomes and verified it as a functional gene in fission yeast. Here we show that 3'-5' helicase activity is intrinsically associated with Hrq1 of Saccharomyces cerevisiae. We also determined several biochemical properties of Hrq1 helicase distinguishable from those of other RecQ helicase members. Hrq1 is able to unwind relatively long duplex DNA up to 120-bp and is significantly stimulated by a preexisting fork structure. Further, the most striking feature of Hrq1 is its absolute requirement for a long 3'-tail (⩾70-nt) for efficient unwinding of duplex DNA. We also found that Hrq1 has potent DNA strand annealing activity. Our results indicate that Hrq1 has vigorous helicase activity that deserves further characterization to expand our understanding of RecQ helicases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Single-Stranded / chemistry*
  • Nucleic Acid Conformation
  • RecQ Helicases / chemistry*
  • RecQ Helicases / genetics
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / chemistry*
  • Saccharomyces cerevisiae Proteins / genetics
  • Substrate Specificity

Substances

  • DNA, Single-Stranded
  • Saccharomyces cerevisiae Proteins
  • Hrq1 protein, S cerevisiae
  • RecQ Helicases