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Reversible fold-switching controls the functional cycle of the antitermination factor RfaH.


ABSTRACT: RfaH, member of the NusG/Spt5 family, activates virulence genes in Gram-negative pathogens. RfaH exists in two states, with its C-terminal domain (CTD) folded either as ?-helical hairpin or ?-barrel. In free RfaH, the ?-helical CTD interacts with, and masks the RNA polymerase binding site on, the N-terminal domain, autoinhibiting RfaH and restricting its recruitment to opsDNA sequences. Upon activation, the domains separate and the CTD refolds into the ?-barrel, which recruits a ribosome, activating translation. Using NMR spectroscopy, we show that only a complete ops-paused transcription elongation complex activates RfaH, probably via a transient encounter complex, allowing the refolded CTD to bind ribosomal protein S10. We also demonstrate that upon release from the elongation complex, the CTD transforms back into the autoinhibitory ?-state, resetting the cycle. Transformation-coupled autoinhibition allows RfaH to achieve high specificity and potent activation of gene expression.

SUBMITTER: Zuber PK 

PROVIDER: S-EPMC6370827 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature

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Reversible fold-switching controls the functional cycle of the antitermination factor RfaH.

Zuber Philipp Konrad PK   Schweimer Kristian K   Rösch Paul P   Artsimovitch Irina I   Knauer Stefan H SH  

Nature communications 20190211 1


RfaH, member of the NusG/Spt5 family, activates virulence genes in Gram-negative pathogens. RfaH exists in two states, with its C-terminal domain (CTD) folded either as α-helical hairpin or β-barrel. In free RfaH, the α-helical CTD interacts with, and masks the RNA polymerase binding site on, the N-terminal domain, autoinhibiting RfaH and restricting its recruitment to opsDNA sequences. Upon activation, the domains separate and the CTD refolds into the β-barrel, which recruits a ribosome, activa  ...[more]

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