Assembly principles of the human R2TP chaperone complex reveal the presence of R2T and R2P complexes

Thiago V Seraphim¹, Nardin Nano², Yiu Wing Sunny Cheung², Siripat Aluksanasuwan³, Carolina Colleti⁴, Yu-Qian Mao², Vaibhav Bhandari², Gavin Young⁵, Larissa Höll⁶, Sadhna Phanse¹, Yuliya Gordiyenko⁵, Daniel R Southworth⁷, Carol V Robinson⁵, Visith Thongboonkerd⁸, Lisandra M Gava⁹, Júlio C Borges¹⁰, Mohan Babu⁶, Leandro R S Barbosa¹¹, Carlos H I Ramos¹², Philipp Kukura⁵, Walid A Houry¹³

Affiliations

¹ Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada; Department of Chemistry and Biochemistry, University of Regina, Regina, SK S4S 0A2, Canada.
² Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada.
³ Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada; Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
⁴ Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada; Center of Biological and Health Sciences, Federal University of São Carlos, São Carlos, SP 13560-970, Brazil.
⁵ Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, Oxford OX1 3QZ, UK.
⁶ Department of Chemistry and Biochemistry, University of Regina, Regina, SK S4S 0A2, Canada.
⁷ Department of Biochemistry and Biophysics, Institute for Neurodegenerative Diseases, University of California, San Francisco, CA 94158, USA.
⁸ Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
⁹ Center of Biological and Health Sciences, Federal University of São Carlos, São Carlos, SP 13560-970, Brazil.
¹⁰ São Carlos Institute of Chemistry, University of São Paulo, São Carlos, SP 13566-590, Brazil.
¹¹ Institute of Physics, University of São Paulo, São Paulo, SP 05508-090, Brazil; Brazilian Synchrotron Light Laboratory (LNLS), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, SP 13083-100, Brazil.
¹² Institute of Chemistry, University of Campinas (UNICAMP), Campinas, SP 13083-970, Brazil.
¹³ Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada; Department of Chemistry, University of Toronto, Toronto, ON M5S 3H6, Canada. Electronic address: walid.houry@utoronto.ca.

PMID: 34492227
DOI: 10.1016/j.str.2021.08.002

Free article

Assembly principles of the human R2TP chaperone complex reveal the presence of R2T and R2P complexes

Thiago V Seraphim et al. Structure. 2022.

Free article

. 2022 Jan 6;30(1):156-171.e12.

doi: 10.1016/j.str.2021.08.002. Epub 2021 Sep 6.

Authors

Affiliations

¹ Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada; Department of Chemistry and Biochemistry, University of Regina, Regina, SK S4S 0A2, Canada.
² Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada.
³ Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada; Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
⁴ Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada; Center of Biological and Health Sciences, Federal University of São Carlos, São Carlos, SP 13560-970, Brazil.
⁵ Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, Oxford OX1 3QZ, UK.
⁶ Department of Chemistry and Biochemistry, University of Regina, Regina, SK S4S 0A2, Canada.
⁷ Department of Biochemistry and Biophysics, Institute for Neurodegenerative Diseases, University of California, San Francisco, CA 94158, USA.
⁸ Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
⁹ Center of Biological and Health Sciences, Federal University of São Carlos, São Carlos, SP 13560-970, Brazil.
¹⁰ São Carlos Institute of Chemistry, University of São Paulo, São Carlos, SP 13566-590, Brazil.
¹¹ Institute of Physics, University of São Paulo, São Paulo, SP 05508-090, Brazil; Brazilian Synchrotron Light Laboratory (LNLS), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, SP 13083-100, Brazil.
¹² Institute of Chemistry, University of Campinas (UNICAMP), Campinas, SP 13083-970, Brazil.
¹³ Department of Biochemistry, University of Toronto, 661 University Avenue, MaRS Centre, West Tower, Room 1612, Toronto, ON M5G 1M1, Canada; Department of Chemistry, University of Toronto, Toronto, ON M5S 3H6, Canada. Electronic address: walid.houry@utoronto.ca.

PMID: 34492227
DOI: 10.1016/j.str.2021.08.002

Abstract

R2TP is a highly conserved chaperone complex formed by two AAA+ ATPases, RUVBL1 and RUVBL2, that associate with PIH1D1 and RPAP3 proteins. R2TP acts in promoting macromolecular complex formation. Here, we establish the principles of R2TP assembly. Three distinct RUVBL1/2-based complexes are identified: R2TP, RUVBL1/2-RPAP3 (R2T), and RUVBL1/2-PIH1D1 (R2P). Interestingly, we find that PIH1D1 does not bind to RUVBL1/RUVBL2 in R2TP and does not function as a nucleotide exchange factor; instead, RPAP3 is found to be the central subunit coordinating R2TP architecture and linking PIH1D1 and RUVBL1/2. We also report that RPAP3 contains an intrinsically disordered N-terminal domain mediating interactions with substrates whose sequences are primarily enriched for Armadillo repeat domains and other helical-type domains. Our work provides a clear and consistent model of R2TP complex structure and gives important insights into how a chaperone machine concerned with assembly of folded proteins into multisubunit complexes might work.

Keywords: AAA+ proteins; ATPases; PAQosome; R2TP; RUVBL1/2; macromolecular complex assembly; molecular chaperones; protein folding.

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Conflict of interest statement

Declaration of interests The authors declare no competing interests. P.K. is a founder, director, and shareholder in Refeyn Ltd. G.Y. is a founder, consultant, and shareholder in Refeyn Ltd.

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Assembly principles of the human R2TP chaperone complex reveal the presence of R2T and R2P complexes

Affiliations

Assembly principles of the human R2TP chaperone complex reveal the presence of R2T and R2P complexes

Authors

Affiliations

Abstract

Conflict of interest statement

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