A Structural Study of the Cytoplasmic Chaperone Effect of 14-3-3 Proteins on Ataxin-1

Seppe Leysen; Rebecca Jane Burnley; Elizabeth Rodriguez; Lech Gustav Milroy; Lorenzo Soini; Carolyn J. Adamski; Larissa Nitschke; Rachel Davis; Tomas Obsil; Lucas Brunsveld; Tom Crabbe; Huda Yahya Zoghbi; Christian Ottmann; Jeremy Martin Davis

doi:10.1016/j.jmb.2021.167174

A Structural Study of the Cytoplasmic Chaperone Effect of 14-3-3 Proteins on Ataxin-1

Seppe Leysen
, Rebecca Jane Burnley
, Elizabeth Rodriguez
, Lech Gustav Milroy
, Lorenzo Soini
, Carolyn J. Adamski
, Larissa Nitschke
, Rachel Davis
, Tomas Obsil
, Lucas Brunsveld
, Tom Crabbe
, Huda Yahya Zoghbi
, Christian Ottmann
, Jeremy Martin Davis (Corresponding author)

Onderzoeksoutput: Bijdrage aan tijdschrift › Tijdschriftartikel › Academic › peer review

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Expansion of the polyglutamine tract in the N terminus of Ataxin-1 is the main cause of the neurodegenerative disease, spinocerebellar ataxia type 1 (SCA1). However, the C-terminal part of the protein – including its AXH domain and a phosphorylation on residue serine 776 – also plays a crucial role in disease development. This phosphorylation event is known to be crucial for the interaction of Ataxin-1 with the 14-3-3 adaptor proteins and has been shown to indirectly contribute to Ataxin-1 stability. Here we show that 14-3-3 also has a direct anti-aggregation or “chaperone” effect on Ataxin-1. Furthermore, we provide structural and biophysical information revealing how phosphorylated S776 in the intrinsically disordered C terminus of Ataxin-1 mediates the cytoplasmic interaction with 14-3-3 proteins. Based on these findings, we propose that 14-3-3 exerts the observed chaperone effect by interfering with Ataxin-1 dimerization through its AXH domain, reducing further self-association. The chaperone effect is particularly important in the context of SCA1, as it was previously shown that a soluble form of mutant Ataxin-1 is the major driver of pathology.

Originele taal-2	Engels
Artikelnummer	167174
Aantal pagina's	20
Tijdschrift	Journal of Molecular Biology
Volume	433
Nummer van het tijdschrift	19
DOI's	https://doi.org/10.1016/j.jmb.2021.167174
Status	Gepubliceerd - 17 sep. 2021

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Publisher Copyright:
© 2021 The Authors

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We wish to thank Dr. Claire Pizzey for collection of SAXS data at the Diamond Light Source and Dr. Petr Pompach at The Czech Academy of Sciences for providing cross-linking/MS data. Initial Training Network TASPPI (H2020 Marie Curie Actions of the European Commission, Grant Agreement 675179); and the Netherlands Organization for Scientific Research (NWO) (Gravity Program 024.001.035 and Vici grant 016.150.366). We wish to thank Dr. Claire Pizzey for collection of SAXS data at the Diamond Light Source and Dr. Petr Pompach at The Czech Academy of Sciences for providing cross-linking/MS data. Initial Training Network TASPPI (H2020 Marie Curie Actions of the European Commission, Grant Agreement 675179); and the Netherlands Organization for Scientific Research (NWO) (Gravity Program 024.001.035 and Vici grant 016.150.366). The authors declare that they have no conflicts of interest with the contents of this article. S.L. C.O, L.B. T.C. and J.M.D initiated the project. S.L. purified proteins, prepared protein crystals, solved and processed crystal structures, processed SAXS data and modelled protein structures. R.J.B. performed and analysed HDX-MS experiments. E.R. performed and analysed AUC experiments. L.G.M. synthesized and purified the ATX pS776 peptide. L.S. performed the ITC measurements. C.A. performed the co-IP experiments. L.N. made the stable DAOY cell lines expressing 2Q, 30Q and 82Q Ataxin-1. S.L. wrote the manuscript with essential input and feedback from all other authors.

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10.1016/j.jmb.2021.167174

published versionDefinitieve gepubliceerde versie, 3,16 MBLicentie: CC BY-NC-ND

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Leysen, S., Burnley, R. J., Rodriguez, E., Milroy, L. G., Soini, L., Adamski, C. J., Nitschke, L., Davis, R., Obsil, T., Brunsveld, L., Crabbe, T., Zoghbi, H. Y., Ottmann, C., & Davis, J. M. (2021). A Structural Study of the Cytoplasmic Chaperone Effect of 14-3-3 Proteins on Ataxin-1. Journal of Molecular Biology, 433(19), Artikel 167174. https://doi.org/10.1016/j.jmb.2021.167174

@article{2f47c358ce244192acc646a8b0c76a63,

title = "A Structural Study of the Cytoplasmic Chaperone Effect of 14-3-3 Proteins on Ataxin-1",

abstract = "Expansion of the polyglutamine tract in the N terminus of Ataxin-1 is the main cause of the neurodegenerative disease, spinocerebellar ataxia type 1 (SCA1). However, the C-terminal part of the protein – including its AXH domain and a phosphorylation on residue serine 776 – also plays a crucial role in disease development. This phosphorylation event is known to be crucial for the interaction of Ataxin-1 with the 14-3-3 adaptor proteins and has been shown to indirectly contribute to Ataxin-1 stability. Here we show that 14-3-3 also has a direct anti-aggregation or “chaperone” effect on Ataxin-1. Furthermore, we provide structural and biophysical information revealing how phosphorylated S776 in the intrinsically disordered C terminus of Ataxin-1 mediates the cytoplasmic interaction with 14-3-3 proteins. Based on these findings, we propose that 14-3-3 exerts the observed chaperone effect by interfering with Ataxin-1 dimerization through its AXH domain, reducing further self-association. The chaperone effect is particularly important in the context of SCA1, as it was previously shown that a soluble form of mutant Ataxin-1 is the major driver of pathology.",

keywords = "crystal structure, HDX-MS, neurodegeneration, protein aggregation, SAXS",

author = "Seppe Leysen and Burnley, \{Rebecca Jane\} and Elizabeth Rodriguez and Milroy, \{Lech Gustav\} and Lorenzo Soini and Adamski, \{Carolyn J.\} and Larissa Nitschke and Rachel Davis and Tomas Obsil and Lucas Brunsveld and Tom Crabbe and Zoghbi, \{Huda Yahya\} and Christian Ottmann and Davis, \{Jeremy Martin\}",

note = "Funding Information: We wish to thank Dr. Claire Pizzey for collection of SAXS data at the Diamond Light Source and Dr. Petr Pompach at The Czech Academy of Sciences for providing cross-linking/MS data. Initial Training Network TASPPI (H2020 Marie Curie Actions of the European Commission, Grant Agreement 675179); and the Netherlands Organization for Scientific Research (NWO) (Gravity Program 024.001.035 and Vici grant 016.150.366). Funding Information: We wish to thank Dr. Claire Pizzey for collection of SAXS data at the Diamond Light Source and Dr. Petr Pompach at The Czech Academy of Sciences for providing cross-linking/MS data. Initial Training Network TASPPI (H2020 Marie Curie Actions of the European Commission, Grant Agreement 675179); and the Netherlands Organization for Scientific Research (NWO) (Gravity Program 024.001.035 and Vici grant 016.150.366). The authors declare that they have no conflicts of interest with the contents of this article. S.L. C.O, L.B. T.C. and J.M.D initiated the project. S.L. purified proteins, prepared protein crystals, solved and processed crystal structures, processed SAXS data and modelled protein structures. R.J.B. performed and analysed HDX-MS experiments. E.R. performed and analysed AUC experiments. L.G.M. synthesized and purified the ATX pS776 peptide. L.S. performed the ITC measurements. C.A. performed the co-IP experiments. L.N. made the stable DAOY cell lines expressing 2Q, 30Q and 82Q Ataxin-1. S.L. wrote the manuscript with essential input and feedback from all other authors. Publisher Copyright: {\textcopyright} 2021 The Authors",

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month = sep,

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doi = "10.1016/j.jmb.2021.167174",

language = "English",

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T1 - A Structural Study of the Cytoplasmic Chaperone Effect of 14-3-3 Proteins on Ataxin-1

AU - Leysen, Seppe

AU - Burnley, Rebecca Jane

AU - Rodriguez, Elizabeth

AU - Milroy, Lech Gustav

AU - Soini, Lorenzo

AU - Adamski, Carolyn J.

AU - Nitschke, Larissa

AU - Davis, Rachel

AU - Obsil, Tomas

AU - Brunsveld, Lucas

AU - Crabbe, Tom

AU - Zoghbi, Huda Yahya

AU - Ottmann, Christian

AU - Davis, Jeremy Martin

N1 - Funding Information: We wish to thank Dr. Claire Pizzey for collection of SAXS data at the Diamond Light Source and Dr. Petr Pompach at The Czech Academy of Sciences for providing cross-linking/MS data. Initial Training Network TASPPI (H2020 Marie Curie Actions of the European Commission, Grant Agreement 675179); and the Netherlands Organization for Scientific Research (NWO) (Gravity Program 024.001.035 and Vici grant 016.150.366). Funding Information: We wish to thank Dr. Claire Pizzey for collection of SAXS data at the Diamond Light Source and Dr. Petr Pompach at The Czech Academy of Sciences for providing cross-linking/MS data. Initial Training Network TASPPI (H2020 Marie Curie Actions of the European Commission, Grant Agreement 675179); and the Netherlands Organization for Scientific Research (NWO) (Gravity Program 024.001.035 and Vici grant 016.150.366). The authors declare that they have no conflicts of interest with the contents of this article. S.L. C.O, L.B. T.C. and J.M.D initiated the project. S.L. purified proteins, prepared protein crystals, solved and processed crystal structures, processed SAXS data and modelled protein structures. R.J.B. performed and analysed HDX-MS experiments. E.R. performed and analysed AUC experiments. L.G.M. synthesized and purified the ATX pS776 peptide. L.S. performed the ITC measurements. C.A. performed the co-IP experiments. L.N. made the stable DAOY cell lines expressing 2Q, 30Q and 82Q Ataxin-1. S.L. wrote the manuscript with essential input and feedback from all other authors. Publisher Copyright: © 2021 The Authors

PY - 2021/9/17

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N2 - Expansion of the polyglutamine tract in the N terminus of Ataxin-1 is the main cause of the neurodegenerative disease, spinocerebellar ataxia type 1 (SCA1). However, the C-terminal part of the protein – including its AXH domain and a phosphorylation on residue serine 776 – also plays a crucial role in disease development. This phosphorylation event is known to be crucial for the interaction of Ataxin-1 with the 14-3-3 adaptor proteins and has been shown to indirectly contribute to Ataxin-1 stability. Here we show that 14-3-3 also has a direct anti-aggregation or “chaperone” effect on Ataxin-1. Furthermore, we provide structural and biophysical information revealing how phosphorylated S776 in the intrinsically disordered C terminus of Ataxin-1 mediates the cytoplasmic interaction with 14-3-3 proteins. Based on these findings, we propose that 14-3-3 exerts the observed chaperone effect by interfering with Ataxin-1 dimerization through its AXH domain, reducing further self-association. The chaperone effect is particularly important in the context of SCA1, as it was previously shown that a soluble form of mutant Ataxin-1 is the major driver of pathology.

AB - Expansion of the polyglutamine tract in the N terminus of Ataxin-1 is the main cause of the neurodegenerative disease, spinocerebellar ataxia type 1 (SCA1). However, the C-terminal part of the protein – including its AXH domain and a phosphorylation on residue serine 776 – also plays a crucial role in disease development. This phosphorylation event is known to be crucial for the interaction of Ataxin-1 with the 14-3-3 adaptor proteins and has been shown to indirectly contribute to Ataxin-1 stability. Here we show that 14-3-3 also has a direct anti-aggregation or “chaperone” effect on Ataxin-1. Furthermore, we provide structural and biophysical information revealing how phosphorylated S776 in the intrinsically disordered C terminus of Ataxin-1 mediates the cytoplasmic interaction with 14-3-3 proteins. Based on these findings, we propose that 14-3-3 exerts the observed chaperone effect by interfering with Ataxin-1 dimerization through its AXH domain, reducing further self-association. The chaperone effect is particularly important in the context of SCA1, as it was previously shown that a soluble form of mutant Ataxin-1 is the major driver of pathology.

KW - crystal structure

KW - HDX-MS

KW - neurodegeneration

KW - protein aggregation

KW - SAXS

UR - https://www.scopus.com/pages/publications/85111313081

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VL - 433

JO - Journal of Molecular Biology

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M1 - 167174

ER -

A Structural Study of the Cytoplasmic Chaperone Effect of 14-3-3 Proteins on Ataxin-1

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