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Structure paper

TitleA mechanistic model of primer synthesis from catalytic structures of DNA polymerase α-primase.
Journal, issue, pagesNat Struct Mol Biol, Year 2024
Publish dateMar 15, 2024
AuthorsElwood A Mullins / Lauren E Salay / Clarissa L Durie / Noah P Bradley / Jane E Jackman / Melanie D Ohi / Walter J Chazin / Brandt F Eichman /
PubMed AbstractThe mechanism by which polymerase α-primase (polα-primase) synthesizes chimeric RNA-DNA primers of defined length and composition, necessary for replication fidelity and genome stability, is ...The mechanism by which polymerase α-primase (polα-primase) synthesizes chimeric RNA-DNA primers of defined length and composition, necessary for replication fidelity and genome stability, is unknown. Here, we report cryo-EM structures of Xenopus laevis polα-primase in complex with primed templates representing various stages of DNA synthesis. Our data show how interaction of the primase regulatory subunit with the primer 5' end facilitates handoff of the primer to polα and increases polα processivity, thereby regulating both RNA and DNA composition. The structures detail how flexibility within the heterotetramer enables synthesis across two active sites and provide evidence that termination of DNA synthesis is facilitated by reduction of polα and primase affinities for the varied conformations along the chimeric primer-template duplex. Together, these findings elucidate a critical catalytic step in replication initiation and provide a comprehensive model for primer synthesis by polα-primase.
External linksNat Struct Mol Biol / PubMed:38491139
MethodsEM (single particle)
Resolution2.8 - 14.06 Å
Structure data

EMDB-29862, PDB-8g99:
Partial auto-inhibitory complex of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 2.8 Å

EMDB-29864, PDB-8g9f:
Complete auto-inhibitory complex of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-29871, PDB-8g9l:
DNA initiation subcomplex of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-29872, PDB-8g9n:
Partial DNA elongation subcomplex of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-29873, PDB-8g9o:
Complete DNA elongation subcomplex of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-29888, PDB-8v5m:
Tetramer core subcomplex (conformation 1) of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 5.3 Å

EMDB-29889, PDB-8v5n:
Tetramer core subcomplex (conformation 2) of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-29891, PDB-8v5o:
Tetramer core subcomplex (conformation 3) of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 4.7 Å

EMDB-42140, PDB-8ucu:
Partial DNA termination subcomplex of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 2.85 Å

EMDB-42141, PDB-8ucv:
Complete DNA termination subcomplex 1 of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 3.81 Å

EMDB-42142, PDB-8ucw:
Complete DNA termination subcomplex 2 of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 3.64 Å

EMDB-42990, PDB-8v6g:
DNA initiation complex (configuration 1) of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 11.16 Å

EMDB-42991, PDB-8v6h:
DNA initiation complex (configuration 2) of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 11.11 Å

EMDB-42992, PDB-8v6i:
DNA elongation complex (configuration 1) of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 14.06 Å

EMDB-42993, PDB-8v6j:
DNA elongation complex (configuration 2) of Xenopus laevis DNA polymerase alpha-primase
Method: EM (single particle) / Resolution: 11.11 Å

Chemicals

ChemComp-ZN:
Unknown entry

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

ChemComp-DGT:
2'-DEOXYGUANOSINE-5'-TRIPHOSPHATE / Deoxyguanosine triphosphate

ChemComp-MG:
Unknown entry

Source
  • xenopus laevis (African clawed frog)
  • synthetic construct (others)
KeywordsREPLICATION / TRANSFERASE / Primase / DNA polymerase / chimeric RNA-DNA primer / RNA/DNA hybrid / DNA replication / DNA synthesis / TRANSFERASE/DNA/RNA / TRANSFERASE-DNA-RNA complex

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