8PID

backtracked E. coli transcription complex paused at ops site and bound to RfaH

8PID の概要

• エントリーDOI: 10.2210/pdb8pid/pdb
• 関連するPDBエントリー: 8PDY 8PEN 8PFG 8PFJ 8PH9 8PHK 8PIB 8PIL 8PIM
• EMDBエントリー: 17681
• 分子名称: Transcription antitermination protein RfaH, MAGNESIUM ION, DNA-directed RNA polymerase subunit beta, ... (10 entities in total)
• 機能のキーワード: transcription complex, pausing, backtracking, rfah, transcription
• 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: 440101.65

構造登録者

Zuber, P.K.,Said, N.,Hilal, T.,Loll, B.,Wahl, M.C.,Knauer, S.H. (登録日: 2023-06-21, 公開日: 2024-04-24)

主引用文献

Zuber, P.K.,Said, N.,Hilal, T.,Wang, B.,Loll, B.,Gonzalez-Higueras, J.,Ramirez-Sarmiento, C.A.,Belogurov, G.A.,Artsimovitch, I.,Wahl, M.C.,Knauer, S.H.
Concerted transformation of a hyper-paused transcription complex and its reinforcing protein.
Nat Commun, 15:3040-3040, 2024

PubMed Abstract: RfaH, a paralog of the universally conserved NusG, binds to RNA polymerases (RNAP) and ribosomes to activate expression of virulence genes. In free, autoinhibited RfaH, an α-helical KOW domain sequesters the RNAP-binding site. Upon recruitment to RNAP paused at an ops site, KOW is released and refolds into a β-barrel, which binds the ribosome. Here, we report structures of ops-paused transcription elongation complexes alone and bound to the autoinhibited and activated RfaH, which reveal swiveled, pre-translocated pause states stabilized by an ops hairpin in the non-template DNA. Autoinhibited RfaH binds and twists the ops hairpin, expanding the RNA:DNA hybrid to 11 base pairs and triggering the KOW release. Once activated, RfaH hyper-stabilizes the pause, which thus requires anti-backtracking factors for escape. Our results suggest that the entire RfaH cycle is solely determined by the ops and RfaH sequences and provide insights into mechanisms of recruitment and metamorphosis of NusG homologs across all life.

PubMed: 38589445
DOI: 10.1038/s41467-024-47368-4

実験手法

ELECTRON MICROSCOPY (3 Å)