Yeast Pif1 accelerates annealing of complementary DNA strands

Biochemistry. 2014 Dec 9;53(48):7659-69. doi: 10.1021/bi500746v. Epub 2014 Nov 26.

Abstract

Pif1 is a helicase involved in the maintenance of nuclear and mitochondrial genomes in eukaryotes. Here we report a new activity of Saccharomyces cerevisiae Pif1, annealing of complementary DNA strands. We identified preferred substrates for annealing as those that generate a duplex product with a single-stranded overhang relative to a blunt end duplex. Importantly, we show that Pif1 can anneal DNA in the presence of ATP and Mg(2+). Pif1-mediated annealing also occurs in the presence of single-stranded DNA binding proteins. Additionally, we show that partial duplex substrates with 3'-single-stranded overhangs such as those generated during double-strand break repair can be annealed by Pif1.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Base Sequence
  • DNA Breaks, Double-Stranded
  • DNA Helicases / metabolism*
  • DNA Repair
  • DNA, Complementary / chemistry*
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism*
  • DNA, Fungal / chemistry*
  • DNA, Fungal / genetics
  • DNA, Fungal / metabolism*
  • Kinetics
  • Magnesium / metabolism
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Substrate Specificity

Substances

  • DNA, Complementary
  • DNA, Fungal
  • Saccharomyces cerevisiae Proteins
  • Adenosine Triphosphate
  • PIF1 protein, S cerevisiae
  • DNA Helicases
  • Magnesium