A1 Refereed original research article in a scientific journal
Concerted transformation of a hyper-paused transcription complex and its reinforcing protein
Authors: Zuber Philipp K., Said Nelly, Hilal Tarek, Wang Bing, Loll Bernhard, González-Higueras Jorge, Ramírez-Sarmiento César A., Belogurov Georgiy A., Artsimovitch Irina, Wahl Markus C., Knauer Stefan H.
Publisher: Springer Nature
Publication year: 2024
Journal: Nature Communications
Journal name in source: Nature communications
Journal acronym: Nat Commun
Article number: 3040
Volume: 15
ISSN: 2041-1723
eISSN: 2041-1723
DOI: https://doi.org/10.1038/s41467-024-47368-4
Web address : https://www.nature.com/articles/s41467-024-47368-4
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/387607704
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.
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