RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation

Jordina Guillén-Boixet; Andrii Kopach; Alex S. Holehouse; Sina Wittmann; Marcus Jahnel; Raimund Schlüßler; Kyoohyun Kim; Irmela R.E.A. Trussina; Jie Wang; Daniel Mateju; Ina Poser; Shovamayee Maharana; Martine Ruer-Gruß; Doris Richter; Xiaojie Zhang; Young Tae Chang; Jochen Guck; Alf Honigmann; Julia Mahamid; Anthony A. Hyman; Rohit V. Pappu; Simon Alberti; Titus M. Franzmann

doi:10.1016/j.cell.2020.03.049

RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation

Jordina Guillén-Boixet, Andrii Kopach, Alex S. Holehouse, Sina Wittmann, Marcus Jahnel, Raimund Schlüßler, Kyoohyun Kim, Irmela R.E.A. Trussina, Jie Wang, Daniel Mateju, Ina Poser, Shovamayee Maharana, Martine Ruer-Gruß, Doris Richter, Xiaojie Zhang, Young Tae Chang, Jochen Guck, Alf Honigmann, Julia Mahamid, Anthony A. HymanRohit V. Pappu, Simon Alberti, Titus M. Franzmann

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

197 Scopus citations

Abstract

Reconstitution of stress granule assembly reveals an autoinhibitory conformation of G3BP that is alleviated by RNA binding, demonstrating how this central node of the stress granule network phase-separates in response to rising cellular RNA concentrations.

Original language	English
Pages (from-to)	346-361.e17
Journal	Cell
Volume	181
Issue number	2
DOIs	https://doi.org/10.1016/j.cell.2020.03.049
State	Published - Apr 16 2020

Keywords

G3BP
Neurodegenerative disease
RNP granules
liquid-to-solid transition
phase separation
stress granules
stress response

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10.1016/j.cell.2020.03.049

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Guillén-Boixet, J., Kopach, A., Holehouse, A. S., Wittmann, S., Jahnel, M., Schlüßler, R., Kim, K., Trussina, I. R. E. A., Wang, J., Mateju, D., Poser, I., Maharana, S., Ruer-Gruß, M., Richter, D., Zhang, X., Chang, Y. T., Guck, J., Honigmann, A., Mahamid, J., ... Franzmann, T. M. (2020). RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation. Cell, 181(2), 346-361.e17. https://doi.org/10.1016/j.cell.2020.03.049

Guillén-Boixet, Jordina ; Kopach, Andrii ; Holehouse, Alex S. ; Wittmann, Sina ; Jahnel, Marcus ; Schlüßler, Raimund ; Kim, Kyoohyun ; Trussina, Irmela R.E.A. ; Wang, Jie ; Mateju, Daniel ; Poser, Ina ; Maharana, Shovamayee ; Ruer-Gruß, Martine ; Richter, Doris ; Zhang, Xiaojie ; Chang, Young Tae ; Guck, Jochen ; Honigmann, Alf ; Mahamid, Julia ; Hyman, Anthony A. ; Pappu, Rohit V. ; Alberti, Simon ; Franzmann, Titus M. / RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation. In: Cell. 2020 ; Vol. 181, No. 2. pp. 346-361.e17.

@article{2478e1f8db1d4c1da81d5badb77b89e0,

title = "RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation",

abstract = "Reconstitution of stress granule assembly reveals an autoinhibitory conformation of G3BP that is alleviated by RNA binding, demonstrating how this central node of the stress granule network phase-separates in response to rising cellular RNA concentrations.",

keywords = "G3BP, Neurodegenerative disease, RNP granules, liquid-to-solid transition, phase separation, stress granules, stress response",

author = "Jordina Guill{\'e}n-Boixet and Andrii Kopach and Holehouse, {Alex S.} and Sina Wittmann and Marcus Jahnel and Raimund Schl{\"u}{\ss}ler and Kyoohyun Kim and Trussina, {Irmela R.E.A.} and Jie Wang and Daniel Mateju and Ina Poser and Shovamayee Maharana and Martine Ruer-Gru{\ss} and Doris Richter and Xiaojie Zhang and Chang, {Young Tae} and Jochen Guck and Alf Honigmann and Julia Mahamid and Hyman, {Anthony A.} and Pappu, {Rohit V.} and Simon Alberti and Franzmann, {Titus M.}",

note = "Funding Information: We thank the Services and Facilities of the MPI-CBG for their support: Protein Expression Purification and Characterization, Light Microscopy, Advanced Imaging, Technology Development Studio, and Scientific Computing Facility. We thank the MPI-CBG staff for helpful discussions, A. Pozniakovsky for help with cloning, and N. Kedersha for wild-type and G3BP1/2 KO U2OS cell lines. We acknowledge funding by the Max-Planck Society; the Technische Universit?t Dresden; the MaxSynBio Consortium; the European Research Council (PhaseAge, 725836); the Bundesministerium f?r Bildung und Forschung (BMBF; 01ED1601A and 031A359A); the Volkswagen Foundation (92847); EU Joint Program ? Neurodegenerative Disease Research (JPND; CureALS); and the European Union Horizon 2020 Research and Innovation Program (Marie Sklodowska-Curie grant agreement 791147). Further support was from the Human Frontier Science Program (RGP0034/2017 to S.A. and R.V.P.), the National Science Foundation (MCB-1614766 to R.V.P.), and St. Jude Children's Research Hospital (to R.V.P.). J.G.-B. and S.W. were supported by EMBO long-term fellowships (ALTF 406-2017 and 708-2017). This paper is the result of the PhD project of A.K. who performed experiments under the supervision of S.A. and T.M.F, and a postdoctoral project led by J. G.-B. Analysis and interpretation of the data were performed mainly by A.S.H. R.V.P. T.M.F. J.G.-B. and S.A. S.M. helped with RNA-binding experiments. I.P. generated HeLa lines. S.W. and J.W. performed co-partitioning experiments. M.R.-G. helped with protein purification. M.J. helped with and analyzed tweezer experiments. D.R. helped with generating constructs. A.H. helped with FCS experiments. X.Z. helped with condensate acquisition of cryo-EM tomography. J.M. reconstructed tomograms and helped with data interpretation. Y.-T.C. provided F22 RNA-staining dye. D.M. helped with translation assays in U2OS cells. I.R.E.A.T assisted with RNA aggregation experiments. R.S. K.K. and J.G. assisted with optical diffraction tomography measurements. A.S.H. and R.V.P. performed simulations and generated predictions regarding the overall phase behavior of G3BP. T.M.F. J.G.-B. and S.A. conceived the project. T.M.F. A.K. J.G.-B. and S.A. drafted the manuscript. R.V.P. A.S.H. T.M.F. J.G.-B. A.A.H. and S.A. worked together to write and edit the manuscript. Funding was acquired by R.V.P. A.A.H. and S.A. S.A. and R.V.P. are advisors on the scientific advisory board of Dewpoint Therapeutics. A.A.H. is a co-founder of Dewpoint Therapeutics. Funding Information: We thank the Services and Facilities of the MPI-CBG for their support: Protein Expression Purification and Characterization, Light Microscopy, Advanced Imaging, Technology Development Studio, and Scientific Computing Facility. We thank the MPI-CBG staff for helpful discussions, A. Pozniakovsky for help with cloning, and N. Kedersha for wild-type and G3BP1/2 KO U2OS cell lines. We acknowledge funding by the Max-Planck Society ; the Technische Universit{\"a}t Dresden ; the MaxSynBio Consortium ; the European Research Council (PhaseAge, 725836 ); the Bundesministerium f{\"u}r Bildung und Forschung (BMBF; 01ED1601A and 031A359A ); the Volkswagen Foundation ( 92847 ); EU Joint Program – Neurodegenerative Disease Research (JPND; CureALS); and the European Union Horizon 2020 Research and Innovation Program (Marie Sklodowska-Curie grant agreement 791147 ). Further support was from the Human Frontier Science Program ( RGP0034/2017 to S.A. and R.V.P.), the National Science Foundation ( MCB-1614766 to R.V.P.), and St. Jude Children's Research Hospital (to R.V.P.). J.G.-B. and S.W. were supported by EMBO long-term fellowships ( ALTF 406-2017 and 708-2017 ). Publisher Copyright: {\textcopyright} 2020 The Author(s)",

year = "2020",

month = apr,

day = "16",

doi = "10.1016/j.cell.2020.03.049",

language = "English",

volume = "181",

pages = "346--361.e17",

journal = "Cell",

issn = "0092-8674",

number = "2",

}

Guillén-Boixet, J, Kopach, A, Holehouse, AS, Wittmann, S, Jahnel, M, Schlüßler, R, Kim, K, Trussina, IREA, Wang, J, Mateju, D, Poser, I, Maharana, S, Ruer-Gruß, M, Richter, D, Zhang, X, Chang, YT, Guck, J, Honigmann, A, Mahamid, J, Hyman, AA, Pappu, RV, Alberti, S & Franzmann, TM 2020, 'RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation', Cell, vol. 181, no. 2, pp. 346-361.e17. https://doi.org/10.1016/j.cell.2020.03.049

RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation. / Guillén-Boixet, Jordina; Kopach, Andrii; Holehouse, Alex S.; Wittmann, Sina; Jahnel, Marcus; Schlüßler, Raimund; Kim, Kyoohyun; Trussina, Irmela R.E.A.; Wang, Jie; Mateju, Daniel; Poser, Ina; Maharana, Shovamayee; Ruer-Gruß, Martine; Richter, Doris; Zhang, Xiaojie; Chang, Young Tae; Guck, Jochen; Honigmann, Alf; Mahamid, Julia; Hyman, Anthony A.; Pappu, Rohit V.; Alberti, Simon; Franzmann, Titus M.

In: Cell, Vol. 181, No. 2, 16.04.2020, p. 346-361.e17.

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

TY - JOUR

T1 - RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation

AU - Guillén-Boixet, Jordina

AU - Kopach, Andrii

AU - Holehouse, Alex S.

AU - Wittmann, Sina

AU - Jahnel, Marcus

AU - Schlüßler, Raimund

AU - Kim, Kyoohyun

AU - Trussina, Irmela R.E.A.

AU - Wang, Jie

AU - Mateju, Daniel

AU - Poser, Ina

AU - Maharana, Shovamayee

AU - Ruer-Gruß, Martine

AU - Richter, Doris

AU - Zhang, Xiaojie

AU - Chang, Young Tae

AU - Guck, Jochen

AU - Honigmann, Alf

AU - Mahamid, Julia

AU - Hyman, Anthony A.

AU - Pappu, Rohit V.

AU - Alberti, Simon

AU - Franzmann, Titus M.

N1 - Funding Information: We thank the Services and Facilities of the MPI-CBG for their support: Protein Expression Purification and Characterization, Light Microscopy, Advanced Imaging, Technology Development Studio, and Scientific Computing Facility. We thank the MPI-CBG staff for helpful discussions, A. Pozniakovsky for help with cloning, and N. Kedersha for wild-type and G3BP1/2 KO U2OS cell lines. We acknowledge funding by the Max-Planck Society; the Technische Universit?t Dresden; the MaxSynBio Consortium; the European Research Council (PhaseAge, 725836); the Bundesministerium f?r Bildung und Forschung (BMBF; 01ED1601A and 031A359A); the Volkswagen Foundation (92847); EU Joint Program ? Neurodegenerative Disease Research (JPND; CureALS); and the European Union Horizon 2020 Research and Innovation Program (Marie Sklodowska-Curie grant agreement 791147). Further support was from the Human Frontier Science Program (RGP0034/2017 to S.A. and R.V.P.), the National Science Foundation (MCB-1614766 to R.V.P.), and St. Jude Children's Research Hospital (to R.V.P.). J.G.-B. and S.W. were supported by EMBO long-term fellowships (ALTF 406-2017 and 708-2017). This paper is the result of the PhD project of A.K. who performed experiments under the supervision of S.A. and T.M.F, and a postdoctoral project led by J. G.-B. Analysis and interpretation of the data were performed mainly by A.S.H. R.V.P. T.M.F. J.G.-B. and S.A. S.M. helped with RNA-binding experiments. I.P. generated HeLa lines. S.W. and J.W. performed co-partitioning experiments. M.R.-G. helped with protein purification. M.J. helped with and analyzed tweezer experiments. D.R. helped with generating constructs. A.H. helped with FCS experiments. X.Z. helped with condensate acquisition of cryo-EM tomography. J.M. reconstructed tomograms and helped with data interpretation. Y.-T.C. provided F22 RNA-staining dye. D.M. helped with translation assays in U2OS cells. I.R.E.A.T assisted with RNA aggregation experiments. R.S. K.K. and J.G. assisted with optical diffraction tomography measurements. A.S.H. and R.V.P. performed simulations and generated predictions regarding the overall phase behavior of G3BP. T.M.F. J.G.-B. and S.A. conceived the project. T.M.F. A.K. J.G.-B. and S.A. drafted the manuscript. R.V.P. A.S.H. T.M.F. J.G.-B. A.A.H. and S.A. worked together to write and edit the manuscript. Funding was acquired by R.V.P. A.A.H. and S.A. S.A. and R.V.P. are advisors on the scientific advisory board of Dewpoint Therapeutics. A.A.H. is a co-founder of Dewpoint Therapeutics. Funding Information: We thank the Services and Facilities of the MPI-CBG for their support: Protein Expression Purification and Characterization, Light Microscopy, Advanced Imaging, Technology Development Studio, and Scientific Computing Facility. We thank the MPI-CBG staff for helpful discussions, A. Pozniakovsky for help with cloning, and N. Kedersha for wild-type and G3BP1/2 KO U2OS cell lines. We acknowledge funding by the Max-Planck Society ; the Technische Universität Dresden ; the MaxSynBio Consortium ; the European Research Council (PhaseAge, 725836 ); the Bundesministerium für Bildung und Forschung (BMBF; 01ED1601A and 031A359A ); the Volkswagen Foundation ( 92847 ); EU Joint Program – Neurodegenerative Disease Research (JPND; CureALS); and the European Union Horizon 2020 Research and Innovation Program (Marie Sklodowska-Curie grant agreement 791147 ). Further support was from the Human Frontier Science Program ( RGP0034/2017 to S.A. and R.V.P.), the National Science Foundation ( MCB-1614766 to R.V.P.), and St. Jude Children's Research Hospital (to R.V.P.). J.G.-B. and S.W. were supported by EMBO long-term fellowships ( ALTF 406-2017 and 708-2017 ). Publisher Copyright: © 2020 The Author(s)

PY - 2020/4/16

Y1 - 2020/4/16

N2 - Reconstitution of stress granule assembly reveals an autoinhibitory conformation of G3BP that is alleviated by RNA binding, demonstrating how this central node of the stress granule network phase-separates in response to rising cellular RNA concentrations.

AB - Reconstitution of stress granule assembly reveals an autoinhibitory conformation of G3BP that is alleviated by RNA binding, demonstrating how this central node of the stress granule network phase-separates in response to rising cellular RNA concentrations.

KW - G3BP

KW - Neurodegenerative disease

KW - RNP granules

KW - liquid-to-solid transition

KW - phase separation

KW - stress granules

KW - stress response

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U2 - 10.1016/j.cell.2020.03.049

DO - 10.1016/j.cell.2020.03.049

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AN - SCOPUS:85083031389

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SP - 346-361.e17

JO - Cell

JF - Cell

SN - 0092-8674

IS - 2

ER -

RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation

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Keywords

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