Human Pif1 helicase unwinds synthetic DNA structures resembling stalled DNA replication forks.
Pif-1 proteins are 5'-->3' superfamily 1 (SF1) helicases that in yeast have roles in the maintenance of mitochondrial and nuclear genome stability. The functions and activities of the human enzyme (hPif1) are unclear, but here we describe its DNA binding and DNA remodeling activities. We demonstrate that hPif1 specifically recognizes and unwinds DNA structures resembling putative stalled replication forks. Notably, the enzyme requires both arms of the replication fork-like structure to initiate efficient unwinding of the putative leading replication strand of such substrates. This DNA structure-specific mode of initiation of unwinding is intrinsic to the conserved core helicase domain (hPifHD) that also possesses a strand annealing activity as has been demonstrated for the RecQ family of helicases. The result of hPif1 helicase action at stalled DNA replication forks would generate free 3' ends and ssDNA that could potentially be used to assist replication restart in conjunction with its strand annealing activity.
George
Tresa
T
Institute for Cancer Studies, University of Sheffield, Beech Hill Road, Sheffield S10 2RX, UK.
Wen
Qin
Q
Griffiths
Richard
R
Ganesh
Anil
A
Meuth
Mark
M
Sanders
Cyril M
CM
D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004265:DNA Helicases / Q000737:chemistry / Q000378:metabolism*; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000378:metabolism; D006801:Humans; D017434:Protein Structure, Tertiary
D004277:DNA, Single-Stranded; D004247:DNA; D004265:DNA Helicases; C508765:PIF1 protein, human
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