Physical Basis for the Loading of a Bacterial Replicative Helicase onto DNA

Ernesto Arias-Palomo, Neha Puri, Valerie L. O'Shea Murray, Qianyun Yan, James M. Berger

Research output: Contribution to journalArticlepeer-review

11 Scopus citations


In cells, dedicated AAA+ ATPases deposit hexameric, ring-shaped helicases onto DNA to initiate chromosomal replication. To better understand the mechanisms by which helicase loading can occur, we used cryo-EM to determine sub-4-Å-resolution structures of the E. coli DnaB⋅DnaC helicase⋅loader complex with nucleotide in pre- and post-DNA engagement states. In the absence of DNA, six DnaC protomers latch onto and crack open a DnaB hexamer using an extended N-terminal domain, stabilizing this conformation through nucleotide-dependent ATPase interactions. Upon binding DNA, DnaC hydrolyzes ATP, allowing DnaB to isomerize into a topologically closed, pre-translocation state competent to bind primase. Our data show how DnaC opens the DnaB ring and represses the helicase prior to DNA binding and how DnaC ATPase activity is reciprocally regulated by DnaB and DNA. Comparative analyses reveal how the helicase loading mechanism of DnaC parallels and diverges from homologous AAA+ systems involved in DNA replication and transposition. Arias-Palomo et al. present the cryo-EM structures of a replicative bacterial helicase-loader complex (E. coli DnaBC) in pre- and post-loading states, revealing how the loader breaks the helicase ring to deposit it at the origin of replication and how ssDNA engagement closes and activates the helicase.

Original languageEnglish (US)
Pages (from-to)173-184.e4
JournalMolecular cell
Issue number1
StatePublished - Apr 4 2019

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology


Dive into the research topics of 'Physical Basis for the Loading of a Bacterial Replicative Helicase onto DNA'. Together they form a unique fingerprint.

Cite this