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

Thiago V Seraphim; Nardin Nano; Yiu Wing Sunny Cheung; Siripat Aluksanasuwan; Carolina Colleti; Yu-Qian Mao; Vaibhav Bhandari; Gavin Young; Larissa Höll; Sadhna Phanse; Yuliya Gordiyenko; Daniel R Southworth; Carol V Robinson; Visith Thongboonkerd; Lisandra M Gava; Júlio C Borges; Mohan Babu; Leandro R S Barbosa; Carlos H I Ramos; Philipp Kukura; Walid A Houry

doi:10.1016/j.str.2021.08.002

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

Structure. 2022 Jan 6;30(1):156-171.e12. doi: 10.1016/j.str.2021.08.002. Epub 2021 Sep 6.

Authors

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

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ATPases Associated with Diverse Cellular Activities / chemistry
ATPases Associated with Diverse Cellular Activities / metabolism*
Apoptosis Regulatory Proteins / chemistry
Apoptosis Regulatory Proteins / metabolism*
Binding Sites
Carrier Proteins / chemistry
Carrier Proteins / metabolism*
Chromatography, Gel
DNA Helicases / chemistry
DNA Helicases / metabolism*
Humans
Models, Molecular
Multiprotein Complexes / chemistry*
Multiprotein Complexes / metabolism
Protein Conformation
Protein Domains
Protein Structure, Quaternary

Substances

Apoptosis Regulatory Proteins
Carrier Proteins
Multiprotein Complexes
PIH1D1 protein, human
RPAP3 protein, human
ATPases Associated with Diverse Cellular Activities
DNA Helicases
RUVBL1 protein, human
RUVBL2 protein, human