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TitleThe DNA replication initiation protein DnaD recognises a specific strand of the Bacillus subtilis chromosome origin.
Journal, issue, pagesNucleic Acids Res, Vol. 51, Issue 9, Page 4322-4340, Year 2023
Publish dateApr 24, 2023
AuthorsCharles Winterhalter / Simone Pelliciari / Daniel Stevens / Stepan Fenyk / Elie Marchand / Nora B Cronin / Panos Soultanas / Tiago R D Costa / Aravindan Ilangovan / Heath Murray /
PubMed AbstractGenome replication is a fundamental biological activity shared by all organisms. Chromosomal replication proceeds bidirectionally from origins, requiring the loading of two helicases, one for each ...Genome replication is a fundamental biological activity shared by all organisms. Chromosomal replication proceeds bidirectionally from origins, requiring the loading of two helicases, one for each replisome. However, the molecular mechanisms underpinning helicase loading at bacterial chromosome origins (oriC) are unclear. Here we investigated the essential DNA replication initiation protein DnaD in the model organism Bacillus subtilis. A set of DnaD residues required for ssDNA binding was identified, and photo-crosslinking revealed that this ssDNA binding region interacts preferentially with one strand of oriC. Biochemical and genetic data support the model that DnaD recognizes a new single-stranded DNA (ssDNA) motif located in oriC, the DnaD Recognition Element (DRE). Considered with single particle cryo-electron microscopy (cryo-EM) imaging of DnaD, we propose that the location of the DRE within oriC orchestrates strand-specific recruitment of helicase during DNA replication initiation. These findings significantly advance our mechanistic understanding of bidirectional replication from a bacterial chromosome origin.
External linksNucleic Acids Res / PubMed:37093985 / PubMed Central
MethodsEM (single particle)
Resolution5.47 Å
Structure data

16914

8ojj

EMDB-16914, PDB-8ojj:
Cryo-EM structure of the DnaD-NTD tetramer
Method: EM (single particle) / Resolution: 5.47 Å

Source
  • Bacillus subtilis (bacteria)
  • bacillus subtilis subsp. subtilis str. 168 (bacteria)
KeywordsDNA BINDING PROTEIN / Replication helicase loading / small cryo-EM structure

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