Human Pif1 helicase is a G-quadruplex DNA-binding protein with G-quadruplex DNA-unwinding activity.
Pif1 proteins are helicases that in yeast are implicated in the maintenance of genome stability. One activity of Saccharomyces cerevisiae Pif1 is to stabilize DNA sequences that could otherwise form deleterious G4 (G-quadruplex) structures by acting as a G4 resolvase. The present study shows that human Pif1 (hPif1, nuclear form) is a G4 DNA-binding and resolvase protein and that these activities are properties of the conserved helicase domain (amino acids 206-620 of 641, hPifHD). hPif1 preferentially bound synthetic G4 DNA relative to ssDNA (single-stranded DNA), dsDNA (double-stranded DNA) and a partially single-stranded duplex DNA helicase substrate. G4 DNA unwinding, but not binding, required an extended (>10 nucleotide) 5' ssDNA tail, and in competition assays, G4 DNA was an ineffective suppressor of helicase activity compared with ssDNA. These results suggest a distinction between the determinants of G4 DNA binding and the ssDNA interactions required for helicase action and that hPif1 may act on G4 substrates by binding alone or as a resolvase. Human Pif1 could therefore have a role in processing G4 structures that arise in the single-stranded nucleic acid intermediates formed during DNA replication and gene expression.
Institute for Cancer Studies, University of Sheffield, UK. c.m.sanders@sheffield.ac.uk
Cyril M
CM
Sanders
D002467:Cell Nucleus / Q000201:enzymology; D004247:DNA / Q000737:chemistry*; D004265:DNA Helicases / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry*; D054856:G-Quadruplexes; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D007527:Isoenzymes; D011994:Recombinant Proteins; D004247:DNA; D004265:DNA Helicases; C508765:PIF1 protein, human