Molecular choreography of primer synthesis by the eukaryotic Pol ?-primase.

The eukaryotic polymerase ? (Pol ?) synthesizes an RNA-DNA hybrid primer of 20-30 nucleotides. Pol ? 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 ? 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 ? 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.
Mesh Terms:
DNA, DNA Primase, DNA Primers, DNA Replication, DNA-Directed DNA Polymerase, RNA, Saccharomyces cerevisiae
Nat Commun
Date: Jun. 21, 2023
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