7UWE

CryoEM Structure of E. coli Transcription-Coupled Ribonucleotide Excision Repair (TC-RER) complex

7UWE の概要

• エントリーDOI: 10.2210/pdb7uwe/pdb
• EMDBエントリー: 26830 26832
• 分子名称: DNA (29-MER), ZINC ION, RNA (18-MER), ... (10 entities in total)
• 機能のキーワード: transcription-coupled rer, transcription, transferase-hydrolase-dna-rna complex, transferase/hydrolase/dna/rna
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 9
• 化学式量合計: 434781.62

構造登録者

Hao, Z.T.,Grower, M.,Bharati, B.,Proshkin, S.,Epshtein, V.,Svetlov, V.,Nudler, E.,Shamovsky, I. (登録日: 2022-05-03, 公開日: 2023-05-31, 最終更新日: 2024-06-12)

主引用文献

Hao, Z.,Gowder, M.,Proshkin, S.,Bharati, B.K.,Epshtein, V.,Svetlov, V.,Shamovsky, I.,Nudler, E.
RNA polymerase drives ribonucleotide excision DNA repair in E. coli.
Cell, 186:2425-, 2023

PubMed Abstract: Ribonuclease HII (RNaseHII) is the principal enzyme that removes misincorporated ribonucleoside monophosphates (rNMPs) from genomic DNA. Here, we present structural, biochemical, and genetic evidence demonstrating that ribonucleotide excision repair (RER) is directly coupled to transcription. Affinity pull-downs and mass-spectrometry-assisted mapping of in cellulo inter-protein cross-linking reveal the majority of RNaseHII molecules interacting with RNA polymerase (RNAP) in E. coli. Cryoelectron microscopy structures of RNaseHII bound to RNAP during elongation, with and without the target rNMP substrate, show specific protein-protein interactions that define the transcription-coupled RER (TC-RER) complex in engaged and unengaged states. The weakening of RNAP-RNaseHII interactions compromises RER in vivo. The structure-functional data support a model where RNaseHII scans DNA in one dimension in search for rNMPs while "riding" the RNAP. We further demonstrate that TC-RER accounts for a significant fraction of repair events, thereby establishing RNAP as a surveillance "vehicle" for detecting the most frequently occurring replication errors.

PubMed: 37196657
DOI: 10.1016/j.cell.2023.04.029

実験手法

ELECTRON MICROSCOPY (2.9 Å)