Concerted transformation of a hyper-paused transcription complex and its reinforcing protein.


Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
08 Apr 2024
Historique:
received: 22 08 2023
accepted: 28 03 2024
medline: 9 4 2024
pubmed: 9 4 2024
entrez: 8 4 2024
Statut: epublish

Résumé

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.

Identifiants

pubmed: 38589445
doi: 10.1038/s41467-024-47368-4
pii: 10.1038/s41467-024-47368-4
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

3040

Subventions

Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : WA 1126/11-1, project number 433623608
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : INST 130/1064-1 FUGG
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : RO 617/21-1
Organisme : Academy of Finland (Suomen Akatemia)
ID : 341962
Organisme : Foundation for the National Institutes of Health (Foundation for the National Institutes of Health, Inc.)
ID : GM067153

Informations de copyright

© 2024. The Author(s).

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Auteurs

Philipp K Zuber (PK)

Biochemistry IV-Biophysical Chemistry, Universität Bayreuth, Bayreuth, Germany.
MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge, UK.

Nelly Said (N)

Institute of Chemistry and Biochemistry, Laboratory of Structural Biochemistry, Freie Universität Berlin, Berlin, Germany.

Tarek Hilal (T)

Institute of Chemistry and Biochemistry, Laboratory of Structural Biochemistry, Freie Universität Berlin, Berlin, Germany.
Research Center of Electron Microscopy and Core Facility BioSupraMol, Freie Universität Berlin, Berlin, Germany.

Bing Wang (B)

Department of Microbiology and Center for RNA Biology, The Ohio State University, Columbus, OH, USA.

Bernhard Loll (B)

Institute of Chemistry and Biochemistry, Laboratory of Structural Biochemistry, Freie Universität Berlin, Berlin, Germany.

Jorge González-Higueras (J)

Institute for Biological and Medical Engineering, Schools of Engineering, Medicine and Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile.
ANID, Millennium Science Initiative Program, Millennium Institute for Integrative Biology, Santiago, Chile.

César A Ramírez-Sarmiento (CA)

Institute for Biological and Medical Engineering, Schools of Engineering, Medicine and Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile.
ANID, Millennium Science Initiative Program, Millennium Institute for Integrative Biology, Santiago, Chile.

Georgiy A Belogurov (GA)

Department of Life Technologies, University of Turku, Turku, Finland.

Irina Artsimovitch (I)

Department of Microbiology and Center for RNA Biology, The Ohio State University, Columbus, OH, USA. artsimovitch.1@osu.edu.

Markus C Wahl (MC)

Institute of Chemistry and Biochemistry, Laboratory of Structural Biochemistry, Freie Universität Berlin, Berlin, Germany. markus.wahl@fu-berlin.de.
Macromolecular Crystallography, Helmholtz-Zentrum Berlin für Materialien und Energie, Berlin, Germany. markus.wahl@fu-berlin.de.

Stefan H Knauer (SH)

Biochemistry IV-Biophysical Chemistry, Universität Bayreuth, Bayreuth, Germany. Stefan.knauer@bms.com.
Bristol-Myers Squibb GmbH & Co. KGaA, Munich, Germany. Stefan.knauer@bms.com.

Classifications MeSH