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Cited by 29Protein Structure and DynamicsRNA and protein synthesis mechanismsHeat shock proteins research

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Structural and thermodynamic analyses of the β-to-α transformation in RfaH reveal principles of fold-switching proteins

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Zuber, Philipp K, Daviter, Tina, Heißmann, Ramona, Persau, Ulrike, Schweimer, Kristian, Knauer, Stefan H·2022·eLife
DOI 10.7554/elife.76630PMC PMC9683785

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Abstract

The two-domain protein RfaH, a paralog of the universally conserved NusG/Spt5 transcription factors, is regulated by autoinhibition coupled to the reversible conformational switch of its 60-residue C-terminal Kyrpides, Ouzounis, Woese (KOW) domain between an α-hairpin and a β-barrel. In contrast, NusG/Spt5-KOW domains only occur in the β-barrel state. To understand the principles underlying the drastic fold switch in RfaH, we elucidated the thermodynamic stability and the structural dynamics of two RfaH- and four NusG/Spt5-KOW domains by combining biophysical and structural biology methods. We find that the RfaH-KOW β-barrel is thermodynamically less stable than that of most NusG/Spt5-KOWs and we show that it is in equilibrium with a globally unfolded species, which, strikingly, contains two helical regions that prime the transition toward the α-hairpin. Our results suggest that transiently structured elements in the unfolded conformation might drive the global folding transition in metamorphic proteins in general.

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Metadata

Authors
Zuber, P. K., Daviter, T., Heißmann, R., Persau, U., Schweimer, K., & Knauer, S. H.
Affiliations
  • University of Bayreuth (DE)
  • Birkbeck, University of London (GB)
Corresponding author
Knauer, Stefan H
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2022-10-18
Volume
11
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.76630
PMC
PMC9683785
OpenAlex
W4306698552
Cited by
29
References
2
Citations by year
2026: 5 · 2025: 8 · 2024: 7 · 2023: 7 · 2022: 2
FWCI
1.96
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • ERef Bayreuth (University of Bayreuth) (submittedVersion)
Funders
European Cooperation in Science and Technology (CA15126)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
Keywords
RfaH, fold switching, autoinhibition, protein thermodynamic stability, protein folding dynamics, beta-barrel, metamorphic proteins, transcription factors, protein conformational switching
MeSH terms
Escherichia coli, Peptide Elongation Factors, Thermodynamics, Transcription Factors, Trans-Activators, Protein Folding, Escherichia coli Proteins

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