Structural and thermodynamic analyses of the β-to-α transformation in RfaH reveal principles of fold-switching proteins
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, Nu...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-10-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/76630 |
| _version_ | 1811317308417638400 |
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| author | Philipp K Zuber Tina Daviter Ramona Heißmann Ulrike Persau Kristian Schweimer Stefan H Knauer |
| author_facet | Philipp K Zuber Tina Daviter Ramona Heißmann Ulrike Persau Kristian Schweimer Stefan H Knauer |
| author_sort | Philipp K Zuber |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T12:04:37Z |
| format | Article |
| id | doaj.art-1f45784865174612a7eaa32eb9925d28 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-13T12:04:37Z |
| publishDate | 2022-10-01 |
| publisher | eLife Sciences Publications Ltd |
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| series | eLife |
| spelling | doaj.art-1f45784865174612a7eaa32eb9925d282022-12-22T02:47:40ZengeLife Sciences Publications LtdeLife2050-084X2022-10-011110.7554/eLife.76630Structural and thermodynamic analyses of the β-to-α transformation in RfaH reveal principles of fold-switching proteinsPhilipp K Zuber0https://orcid.org/0000-0001-5139-3930Tina Daviter1https://orcid.org/0000-0003-2636-5959Ramona Heißmann2Ulrike Persau3Kristian Schweimer4https://orcid.org/0000-0002-3837-8442Stefan H Knauer5https://orcid.org/0000-0002-4143-0694Biochemistry IV – Biophysical Chemistry, University of Bayreuth, Bayreuth, GermanyBirkbeck, University of London, Malet Street, Bloomsbury, London, United KingdomBiochemistry IV – Biophysical Chemistry, University of Bayreuth, Bayreuth, GermanyBiochemistry IV – Biophysical Chemistry, University of Bayreuth, Bayreuth, GermanyBiochemistry IV – Biophysical Chemistry, University of Bayreuth, Bayreuth, GermanyBiochemistry IV – Biophysical Chemistry, University of Bayreuth, Bayreuth, GermanyThe 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.https://elifesciences.org/articles/76630fold-switchingNusG proteinsRfaHmetamorphic proteinsKOW domains |
| spellingShingle | Philipp K Zuber Tina Daviter Ramona Heißmann Ulrike Persau Kristian Schweimer Stefan H Knauer Structural and thermodynamic analyses of the β-to-α transformation in RfaH reveal principles of fold-switching proteins eLife fold-switching NusG proteins RfaH metamorphic proteins KOW domains |
| title | Structural and thermodynamic analyses of the β-to-α transformation in RfaH reveal principles of fold-switching proteins |
| title_full | Structural and thermodynamic analyses of the β-to-α transformation in RfaH reveal principles of fold-switching proteins |
| title_fullStr | Structural and thermodynamic analyses of the β-to-α transformation in RfaH reveal principles of fold-switching proteins |
| title_full_unstemmed | Structural and thermodynamic analyses of the β-to-α transformation in RfaH reveal principles of fold-switching proteins |
| title_short | Structural and thermodynamic analyses of the β-to-α transformation in RfaH reveal principles of fold-switching proteins |
| title_sort | structural and thermodynamic analyses of the β to α transformation in rfah reveal principles of fold switching proteins |
| topic | fold-switching NusG proteins RfaH metamorphic proteins KOW domains |
| url | https://elifesciences.org/articles/76630 |
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