7P9J

Prim-Pol Domain of CRISPR-associated Prim-Pol (CAPP) from Marinitoga sp. 1137 - Primer Initiation Complex

7P9J の概要

• エントリーDOI: 10.2210/pdb7p9j/pdb
• 関連するPDBエントリー: 7NQD 7NQE 7NQF
• 分子名称: TPR_REGION domain-containing protein, Templating strand, COBALT (II) ION, ... (6 entities in total)
• 機能のキーワード: aep archaeo-eukaryotic primase apo prim-pol primase-polymerase primase polymerase, transferase
• 由来する生物種: Marinitoga sp. 1137; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 83111.40

構造登録者

Li, A.W.H.,Doherty, A.J. (登録日: 2021-07-27, 公開日: 2022-05-04, 最終更新日: 2024-01-31)

主引用文献

Li, A.W.H.,Zabrady, K.,Bainbridge, L.J.,Zabrady, M.,Naseem-Khan, S.,Berger, M.B.,Kolesar, P.,Cisneros, G.A.,Doherty, A.J.
Molecular basis for the initiation of DNA primer synthesis.
Nature, 605:767-773, 2022

PubMed Abstract: During the initiation of DNA replication, oligonucleotide primers are synthesized de novo by primases and are subsequently extended by replicative polymerases to complete genome duplication. The primase-polymerase (Prim-Pol) superfamily is a diverse grouping of primases, which includes replicative primases and CRISPR-associated primase-polymerases (CAPPs) involved in adaptive immunity. Although much is known about the activities of these enzymes, the precise mechanism used by primases to initiate primer synthesis has not been elucidated. Here we identify the molecular bases for the initiation of primer synthesis by CAPP and show that this mechanism is also conserved in replicative primases. The crystal structure of a primer initiation complex reveals how the incoming nucleotides are positioned within the active site, adjacent to metal cofactors and paired to the templating single-stranded DNA strand, before synthesis of the first phosphodiester bond. Furthermore, the structure of a Prim-Pol complex with double-stranded DNA shows how the enzyme subsequently extends primers in a processive polymerase mode. The structural and mechanistic studies presented here establish how Prim-Pol proteins instigate primer synthesis, revealing the requisite molecular determinants for primer synthesis within the catalytic domain. This work also establishes that the catalytic domain of Prim-Pol enzymes, including replicative primases, is sufficient to catalyse primer formation.

PubMed: 35508653
DOI: 10.1038/s41586-022-04695-0

実験手法

X-RAY DIFFRACTION (1.9 Å)