8AMF

Cryo-EM structure of the RecA postsynaptic filament from S. pneumoniae

8AMF の概要

• エントリーDOI: 10.2210/pdb8amf/pdb
• EMDBエントリー: 15525
• 分子名称: Protein RecA, DNA, PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER, ... (4 entities in total)
• 機能のキーワード: helical reconstruction, recombinase, streptococcus pneumoniae, double-stranded dna, recombination
• 由来する生物種: Streptococcus pneumoniae; 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 176207.88

構造登録者

Perry, T.N.,Fronzes, R.,Polard, P.,Hertzog, M. (登録日: 2022-08-03, 公開日: 2024-02-21, 最終更新日: 2024-03-06)

主引用文献

Hertzog, M.,Perry, T.N.,Dupaigne, P.,Serres, S.,Morales, V.,Soulet, A.L.,Bell, J.C.,Margeat, E.,Kowalczykowski, S.C.,Le Cam, E.,Fronzes, R.,Polard, P.
Assembly mechanism and cryoEM structure of RecA recombination nucleofilaments from Streptococcus pneumoniae.
Nucleic Acids Res., 51:2800-2817, 2023

PubMed Abstract: RecA-mediated homologous recombination (HR) is a key mechanism for genome maintenance and plasticity in bacteria. It proceeds through RecA assembly into a dynamic filament on ssDNA, the presynaptic filament, which mediates DNA homology search and ordered DNA strand exchange. Here, we combined structural, single molecule and biochemical approaches to characterize the ATP-dependent assembly mechanism of the presynaptic filament of RecA from Streptococcus pneumoniae (SpRecA), in comparison to the Escherichia coli RecA (EcRecA) paradigm. EcRecA polymerization on ssDNA is assisted by the Single-Stranded DNA Binding (SSB) protein, which unwinds ssDNA secondary structures that block EcRecA nucleofilament growth. We report by direct microscopic analysis of SpRecA filamentation on ssDNA that neither of the two paralogous pneumococcal SSBs could assist the extension of SpRecA nucleopolymers. Instead, we found that the conserved RadA helicase promotes SpRecA nucleofilamentation in an ATP-dependent manner. This allowed us to solve the atomic structure of such a long native SpRecA nucleopolymer by cryoEM stabilized with ATPγS. It was found to be equivalent to the crystal structure of the EcRecA filament with a marked difference in how RecA mediates nucleotide orientation in the stretched ssDNA. Then, our results show that SpRecA and EcRecA HR activities are different, in correlation with their distinct ATP-dependent ssDNA binding modes.

PubMed: 36806960
DOI: 10.1093/nar/gkad080

実験手法

ELECTRON MICROSCOPY (3.8 Å)