Characterization of the Helicase Activity and Anti-telomerase Properties of Yeast Pif1p In Vitro
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 wordsYeast Pif1p helicase telomerase oligonucleotide substrate-based radiometric assays