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.
Boulé
Jean-Baptiste
JB
Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
Zakian
Virginia A
VA
D002458:Cell Fractionation / Q000379:methods; D002845:Chromatography / Q000379:methods; D004265:DNA Helicases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D019098:Telomerase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
D009841:Oligonucleotides; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D019098:Telomerase; C085463:PIF1 protein, S cerevisiae; D004265:DNA Helicases