Archaeal Hel308 helicase targets replication forks in vivo and in vitro and unwinds lagging strands.
Mutations in mammalian and Drosophila Hel308 and PolQ paralogues cause genome instability but their helicase functions are mysterious. By in vivo and in vitro analysis, we show that Hel308 from archaea (Hel308a) may act at stalled replication forks. Introducing hel308a into Escherichia coli dnaE strains that conditionally accumulate stalled forks caused synthetic lethality, an effect indistinguishable from E.coli RecQ. Further analysis in vivo indicated that the effect of hel308a is exerted independently of homologous recombination. The minimal biochemical properties of Hel308a protein were the same as human Hel308. We describe how helicase actions of Hel308a at fork structures lead specifically to displacement of lagging strands. The invading strand of D-loops is also targeted. Using archaeal Hel308, we propose models of action for the helicase domain of PolQ, promoting loading of the translesion polymerase domain. We speculate that removal of lagging strands at stalled forks by Hel308 promotes the formation of initiation zones, priming restart of lagging strand synthesis.
Institute of Genetics, School of Biology, University of Nottingham, Queen's Medical Centre Nottingham NG7 2UH, UK.
Colin P
CP
Guy
Edward L
EL
Bolt
D019843:Archaeal Proteins / Q000145:classification / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004265:DNA Helicases / Q000145:classification / Q000378:metabolism*; D004258:DNA Polymerase III / Q000235:genetics; D004261:DNA Replication; D045566:DNA, Cruciform / Q000378:metabolism; D008957:Models, Genetic; D017354:Point Mutation
D019843:Archaeal Proteins; D045566:DNA, Cruciform; D004247:DNA; D004258:DNA Polymerase III; D004265:DNA Helicases; C451406:HELQ protein, human