Molecular choreography of primer synthesis by the eukaryotic Pol α-primase

Zuanning Yuan, Roxana Georgescu, Huilin Li () and Michael E. O’Donnell ()
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Zuanning Yuan: Van Andel Institute
Roxana Georgescu: Rockefeller University
Huilin Li: Van Andel Institute
Michael E. O’Donnell: Rockefeller University

Nature Communications, 2023, vol. 14, issue 1, 1-14

Abstract: Abstract 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.

Date: 2023
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DOI: 10.1038/s41467-023-39441-1

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