A mechanistic model of primer synthesis from catalytic structures of DNA polymerase α-primase

Elwood A Mullins; Lauren E Salay; Clarissa L Durie; Noah P Bradley; Jane E Jackman; Melanie D Ohi; Walter J Chazin; Brandt F Eichman

doi:10.1038/s41594-024-01227-4

A mechanistic model of primer synthesis from catalytic structures of DNA polymerase α-primase

Nat Struct Mol Biol. 2024 Mar 15. doi: 10.1038/s41594-024-01227-4. Online ahead of print.

Authors

Elwood A Mullins^{1

2}, Lauren E Salay^{2

3

4}, Clarissa L Durie^{5

6

7}, Noah P Bradley^{1

2}, Jane E Jackman^{8

9}, Melanie D Ohi^{5

6}, Walter J Chazin^{10

11

12}, Brandt F Eichman^{13

14

15}

Affiliations

¹ Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA.
² Center for Structural Biology, Vanderbilt University, Nashville, TN, USA.
³ Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA.
⁴ Department of Biochemistry, University of Washington, Seattle, WA, USA.
⁵ Department of Cell and Developmental Biology, University of Michigan School of Medicine, Ann Arbor, MI, USA.
⁶ Life Sciences Institute, University of Michigan, Ann Arbor, MI, USA.
⁷ Department of Biochemistry, University of Missouri, Columbia, MO, USA.
⁸ Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, USA.
⁹ Center for RNA Biology, The Ohio State University, Columbus, OH, USA.
¹⁰ Center for Structural Biology, Vanderbilt University, Nashville, TN, USA. walter.j.chazin@vanderbilt.edu.
¹¹ Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA. walter.j.chazin@vanderbilt.edu.
¹² Department of Chemistry, Vanderbilt University, Nashville, TN, USA. walter.j.chazin@vanderbilt.edu.
¹³ Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA. brandt.eichman@vanderbilt.edu.
¹⁴ Center for Structural Biology, Vanderbilt University, Nashville, TN, USA. brandt.eichman@vanderbilt.edu.
¹⁵ Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA. brandt.eichman@vanderbilt.edu.

PMID: 38491139
DOI: 10.1038/s41594-024-01227-4

Abstract

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.

Abstract

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