The permanently chaperone-active small heat shock protein Hsp17 from Caenorhabditis elegans exhibits topological separation of its N-terminal regions

Annika Strauch; Benjamin Rossa; Fabian Köhler; Simon Haeussler; Moritz Mühlhofer; Florian Rührnößl; Caroline Körösy; Yevheniia Bushman; Barbara Conradt; Martin Haslbeck; Sevil Weinkauf; Johannes Buchner

doi:10.1016/j.jbc.2022.102753

The permanently chaperone-active small heat shock protein Hsp17 from Caenorhabditis elegans exhibits topological separation of its N-terminal regions

Annika Strauch, Benjamin Rossa, Fabian Köhler, Simon Haeussler, Moritz Mühlhofer, Florian Rührnößl, Caroline Körösy, Yevheniia Bushman, Barbara Conradt, Martin Haslbeck, Sevil Weinkauf, Johannes Buchner^*

^*Corresponding author for this work

Sub Molecular Biophysics

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Small Heat shock proteins (sHsps) are a family of molecular chaperones that bind nonnative proteins in an ATP-independent manner. Caenorhabditis elegans encodes 16 different sHsps, among them Hsp17, which is evolutionarily distinct from other sHsps in the nematode. The structure and mechanism of Hsp17 and how these may differ from other sHsps remain unclear. Here, we find that Hsp17 has a distinct expression pattern, structural organization, and chaperone function. Consistent with its presence under nonstress conditions, and in contrast to many other sHsps, we determined that Hsp17 is a mono-disperse, permanently active chaperone in vitro, which interacts with hundreds of different C. elegans proteins under physiological conditions. Additionally, our cryo-EM structure of Hsp17 reveals that in the 24-mer complex, 12 N-terminal regions are involved in its chaperone function. These flexible regions are located on the outside of the spherical oligomer, whereas the other 12 N-terminal regions are engaged in stabilizing interactions in its interior. This allows the same region in Hsp17 to perform different functions depending on the topological context. Taken together, our results reveal structural and functional features that further define the structural basis of permanently active sHsps.

Original language	English
Article number	102753
Number of pages	19
Journal	Journal of Biological Chemistry
Volume	299
Issue number	1
DOIs	https://doi.org/10.1016/j.jbc.2022.102753
Publication status	Published - Jan 2023

Bibliographical note

Publisher Copyright:
© 2022 The Authors

Funding

We thank Dragana A. M. Catici for critical reading of the article, Nadin Memar for discussing the C. elegans experiments, and Laura Meier as well as, Mara Pfeifer, Carmen Chen, Florian Rothfischer, and Tetyana Dashivets for their experimental support. We kindly thank Katja Bäuml for operating the MS instrument and our technical assistants for general support in the laboratory. Dietrich Mostert for the help with the MS samples preparation (insoluble and soluble fraction). We gratefully thank Dr Jürgen Plitzko (Max Planck Institute for Biochemistry, Martinsried, Germany) for ongoing support with EM. Moreover, we thank S. Mitani (National BioResource Project, Tokyo, Japan) for hsp-17 −/− (tm5013) strain and the GCG (founded by NIH Office of Research Infrastructure Programs, P40 OD010440) for the N2 wt strain and hsp-16.2 −/− (VC475) strain. The Deutsche Forschungsgemeinschaft (DFG) Sonderforschungsbereich (1035) project A6 (number 201202640 ) as well as CO204/6-1 (to F. K. and S. H.) and CO204/9-1 (to B. C.) funded this work. We thank Dragana A. M. Catici for critical reading of the article, Nadin Memar for discussing the C. elegans experiments, and Laura Meier as well as, Mara Pfeifer, Carmen Chen, Florian Rothfischer, and Tetyana Dashivets for their experimental support. We kindly thank Katja Bäuml for operating the MS instrument and our technical assistants for general support in the laboratory. Dietrich Mostert for the help with the MS samples preparation (insoluble and soluble fraction). We gratefully thank Dr Jürgen Plitzko (Max Planck Institute for Biochemistry, Martinsried, Germany) for ongoing support with EM. Moreover, we thank S. Mitani (National BioResource Project, Tokyo, Japan) for hsp-17−/− (tm5013) strain and the GCG (founded by NIH Office of Research Infrastructure Programs, P40 OD010440) for the N2 wt strain and hsp-16.2−/− (VC475) strain. The Deutsche Forschungsgemeinschaft (DFG) Sonderforschungsbereich (1035) project A6 (number 201202640) as well as CO204/6-1 (to F. K. and S. H.) and CO204/9-1 (to B. C.) funded this work. A. S. M. H. S. W. and J. B. methodology; A. S. B. R. F. K. S. H. M. M. F. R. C. K. Y. B. and B. C. investigation; A. S. B. R. M. M. F. R. and J. B. formal analysis; B. R. data curation; A. S. B. R. and J. B. writing–original draft.

Funders	Funder number
Laura Meier
Max Planck Institute for Biochemistry	tm5013
National Institutes of Health	P40 OD010440, VC475
Deutsche Forschungsgemeinschaft	CO204/9-1, 201202640, CO204/6-1

Keywords

Hsp-17
Hsp17
molecular chaperone
protein aggregation
protein structure
sHsps
small heat shock protein

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1-s2.0-S0021925822011966-mainFinal published version, 4.35 MBLicence: CC BY

Cite this

Strauch, A., Rossa, B., Köhler, F., Haeussler, S., Mühlhofer, M., Rührnößl, F., Körösy, C., Bushman, Y., Conradt, B., Haslbeck, M., Weinkauf, S., & Buchner, J. (2023). The permanently chaperone-active small heat shock protein Hsp17 from Caenorhabditis elegans exhibits topological separation of its N-terminal regions. Journal of Biological Chemistry, 299(1), Article 102753. https://doi.org/10.1016/j.jbc.2022.102753

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title = "The permanently chaperone-active small heat shock protein Hsp17 from Caenorhabditis elegans exhibits topological separation of its N-terminal regions",

abstract = "Small Heat shock proteins (sHsps) are a family of molecular chaperones that bind nonnative proteins in an ATP-independent manner. Caenorhabditis elegans encodes 16 different sHsps, among them Hsp17, which is evolutionarily distinct from other sHsps in the nematode. The structure and mechanism of Hsp17 and how these may differ from other sHsps remain unclear. Here, we find that Hsp17 has a distinct expression pattern, structural organization, and chaperone function. Consistent with its presence under nonstress conditions, and in contrast to many other sHsps, we determined that Hsp17 is a mono-disperse, permanently active chaperone in vitro, which interacts with hundreds of different C. elegans proteins under physiological conditions. Additionally, our cryo-EM structure of Hsp17 reveals that in the 24-mer complex, 12 N-terminal regions are involved in its chaperone function. These flexible regions are located on the outside of the spherical oligomer, whereas the other 12 N-terminal regions are engaged in stabilizing interactions in its interior. This allows the same region in Hsp17 to perform different functions depending on the topological context. Taken together, our results reveal structural and functional features that further define the structural basis of permanently active sHsps.",

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Strauch, A, Rossa, B, Köhler, F, Haeussler, S, Mühlhofer, M, Rührnößl, F, Körösy, C, Bushman, Y, Conradt, B, Haslbeck, M, Weinkauf, S & Buchner, J 2023, 'The permanently chaperone-active small heat shock protein Hsp17 from Caenorhabditis elegans exhibits topological separation of its N-terminal regions', Journal of Biological Chemistry, vol. 299, no. 1, 102753. https://doi.org/10.1016/j.jbc.2022.102753

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T1 - The permanently chaperone-active small heat shock protein Hsp17 from Caenorhabditis elegans exhibits topological separation of its N-terminal regions

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AU - Rossa, Benjamin

AU - Köhler, Fabian

AU - Haeussler, Simon

AU - Mühlhofer, Moritz

AU - Rührnößl, Florian

AU - Körösy, Caroline

AU - Bushman, Yevheniia

AU - Conradt, Barbara

AU - Haslbeck, Martin

AU - Weinkauf, Sevil

AU - Buchner, Johannes

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AB - Small Heat shock proteins (sHsps) are a family of molecular chaperones that bind nonnative proteins in an ATP-independent manner. Caenorhabditis elegans encodes 16 different sHsps, among them Hsp17, which is evolutionarily distinct from other sHsps in the nematode. The structure and mechanism of Hsp17 and how these may differ from other sHsps remain unclear. Here, we find that Hsp17 has a distinct expression pattern, structural organization, and chaperone function. Consistent with its presence under nonstress conditions, and in contrast to many other sHsps, we determined that Hsp17 is a mono-disperse, permanently active chaperone in vitro, which interacts with hundreds of different C. elegans proteins under physiological conditions. Additionally, our cryo-EM structure of Hsp17 reveals that in the 24-mer complex, 12 N-terminal regions are involved in its chaperone function. These flexible regions are located on the outside of the spherical oligomer, whereas the other 12 N-terminal regions are engaged in stabilizing interactions in its interior. This allows the same region in Hsp17 to perform different functions depending on the topological context. Taken together, our results reveal structural and functional features that further define the structural basis of permanently active sHsps.

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ER -

The permanently chaperone-active small heat shock protein Hsp17 from Caenorhabditis elegans exhibits topological separation of its N-terminal regions

Abstract

Bibliographical note

Funding

Keywords

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