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TitleDomain dynamics within the PriA DNA helicase regulate DNA replication restart.
Journal, issue, pagesNucleic Acids Res, Vol. 54, Issue 18, Year 2026
Publish dateSep 22, 2026
AuthorsHaley R Deorio / Alexander T Duckworth / Gavin R Forsythe / Andrew Y Sung / Steve J Sandler / Timothy Grant / James L Keck /
PubMed AbstractPrematurely terminated DNA replication processes in bacteria must be restarted for successful genome duplication. In Escherichia coli, the PriA DNA helicase orchestrates replication restart by ...Prematurely terminated DNA replication processes in bacteria must be restarted for successful genome duplication. In Escherichia coli, the PriA DNA helicase orchestrates replication restart by assembling the PriA/PriB/DnaT preprimosome complex onto abandoned DNA replication forks and reloading the replicative machinery. We show that the structure of the replication fork lagging strand-whether single- or double-stranded-influences the position of the cysteine-rich region in bound PriA (PriACRR). When PriA binds to synthetic replication forks with a single-stranded lagging strand, the PriACRR is found either in a state similar to free PriA or in a rotated position that encircles the lagging strand and allows PriB recruitment. Binding to a synthetic replication fork with a duplex lagging strand neither alters the PriACRR position nor promotes PriB binding, suggesting PriA must unwind duplex lagging-strand DNA to trigger PriACRR movement and subsequent preprimosome formation. PriA variants designed to destabilize the two PriACRR positions differentially affect ATPase activity, helicase function, PriB binding in vitro, and PriA activity in vivo. These results support a regulatory switch model in which the lagging-strand DNA structure modulates the PriACRR position, thereby governing PriA's biochemical and cellular activities.
External linksNucleic Acids Res / PubMed:42770332 / PubMed Central
MethodsEM (single particle)
Resolution3.5 - 3.8 Å
Structure data

EMDB-76387: EcPriA bound to DNA replication fork with dsDNA lagging strand
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-76388: EcPriA bound to DNA replication fork with ssDNA lagging strand (CRR up)
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-76389: EcPriA bound to DNA replication fork with ssDNA lagging strand (CRR down)
Method: EM (single particle) / Resolution: 3.8 Å

Source
  • Escherichia coli K-12 (bacteria)
  • synthetic construct (others)

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