Structural basis of the T4 bacteriophage primosome assembly and primer synthesis.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
20 07 2023
20 07 2023
Historique:
received:
30
09
2022
accepted:
11
07
2023
medline:
24
7
2023
pubmed:
21
7
2023
entrez:
20
7
2023
Statut:
epublish
Résumé
The T4 bacteriophage gp41 helicase and gp61 primase assemble into a primosome to couple DNA unwinding with RNA primer synthesis for DNA replication. How the primosome is assembled and how the primer length is defined are unclear. Here we report a series of cryo-EM structures of T4 primosome assembly intermediates. We show that gp41 alone is an open spiral, and ssDNA binding triggers a large-scale scissor-like conformational change that drives the ring closure and activates the helicase. Helicase activation exposes a cryptic hydrophobic surface to recruit the gp61 primase. The primase binds the helicase in a bipartite mode in which the N-terminal Zn-binding domain and the C-terminal RNA polymerase domain each contain a helicase-interacting motif that bind to separate gp41 N-terminal hairpin dimers, leading to the assembly of one primase on the helicase hexamer. Our study reveals the T4 primosome assembly process and sheds light on the RNA primer synthesis mechanism.
Identifiants
pubmed: 37474605
doi: 10.1038/s41467-023-40106-2
pii: 10.1038/s41467-023-40106-2
pmc: PMC10359460
doi:
Substances chimiques
DNA Primase
EC 2.7.7.-
DNA Helicases
EC 3.6.4.-
DNA Primers
0
DNA, Viral
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
4396Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM013306
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM131754
Pays : United States
Commentaires et corrections
Type : UpdateOf
Informations de copyright
© 2023. The Author(s).
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