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TitleMolecular choreography of primer synthesis by the eukaryotic Pol α-primase.
Journal, issue, pagesNat Commun, Vol. 14, Issue 1, Page 3697, Year 2023
Publish dateJun 21, 2023
AuthorsZuanning Yuan / Roxana Georgescu / Huilin Li / Michael E O'Donnell /
PubMed AbstractThe eukaryotic polymerase α (Pol α) synthesizes an RNA-DNA hybrid primer of 20-30 nucleotides. Pol α is composed of Pol1, Pol12, Primase 1 (Pri1), and Pri2. Pol1 and Pri1 contain the DNA ...The eukaryotic polymerase α (Pol α) synthesizes an RNA-DNA hybrid primer of 20-30 nucleotides. Pol α is composed of Pol1, Pol12, Primase 1 (Pri1), and Pri2. Pol1 and Pri1 contain the DNA polymerase and RNA primase activities, respectively. It has been unclear how Pol α hands over an RNA primer from Pri1 to Pol1 for DNA primer extension, and how the primer length is defined. Here we report the cryo-EM analysis of yeast Pol α in the apo, primer initiation, primer elongation, RNA primer hand-off from Pri1 to Pol1, and DNA extension states, revealing a series of very large movements. We reveal a critical point at which Pol1-core moves to take over the 3'-end of the RNA from Pri1. DNA extension is limited by a spiral motion of Pol1-core. Since both Pri1 and Pol1-core are flexibly attached to a stable platform, primer growth produces stress that limits the primer length.
External linksNat Commun / PubMed:37344454 / PubMed Central
MethodsEM (single particle)
Resolution3.56 - 5.6 Å
Structure data

29345

8foc

EMDB-29345, PDB-8foc:
Cryo-EM structure of S. cerevisiae DNA polymerase alpha-primase in Apo state conformation I
Method: EM (single particle) / Resolution: 3.7 Å

29346

8fod

EMDB-29346, PDB-8fod:
Cryo-EM structure of S. cerevisiae DNA polymerase alpha-primase complex in Apo state conformation II
Method: EM (single particle) / Resolution: 3.8 Å

29347

8foe

EMDB-29347, PDB-8foe:
Cryo-EM structure of S. cerevisiae DNA polymerase alpha-primase complex bound to a template DNA
Method: EM (single particle) / Resolution: 5.6 Å

29349

8foh

EMDB-29349, PDB-8foh:
Cryo-EM structure of S. cerevisiae DNA polymerase alpha-primase complex in the RNA synthesis state
Method: EM (single particle) / Resolution: 4.6 Å

29351

8foj

EMDB-29351, PDB-8foj:
Cryo-EM structure of S. cerevisiae DNA polymerase alpha-primase complex in the post RNA handoff state
Method: EM (single particle) / Resolution: 4.8 Å

29352

8fok

EMDB-29352, PDB-8fok:
Cryo-EM structure of S. cerevisiae DNA polymerase alpha-primase complex in the DNA elongation state
Method: EM (single particle) / Resolution: 3.56 Å

Chemicals

ChemComp-SF4:
IRON/SULFUR CLUSTER / Iron–sulfur cluster

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • synthetic construct (others)
KeywordsDNA BINDING PROTEIN / DNA polymerase / primase / DNA replication

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