Protocol

Helicases

Volume 587 of the series Methods in Molecular Biology pp 359-376

Date:

Characterization of the Helicase Activity and Anti-telomerase Properties of Yeast Pif1p In Vitro

  • Jean-Baptiste BouléAffiliated withDepartment of Molecular Biology, Princeton University
  • , Virginia A. ZakianAffiliated withDepartment of Molecular Biology, Princeton University

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Abstract

Pif1p is the prototype member of a family of helicases that is highly conserved from yeast to humans. In yeast, Pif1p is involved in many aspects of the preservation of genome stability. In particular, Pif1p is involved in the maintenance of mitochondrial DNA and in the direct inhibition of telomerase at telomeres and double-stranded breaks. Here we describe methods to purify Pif1p and study in vitro its enzymatic properties and functional interaction with telomerase.

Key words

Yeast Pif1p helicase telomerase oligonucleotide substrate-based radiometric assays