Cargando…

Structural and functional analysis of the role of the chaperonin CCT in mTOR complex assembly

The mechanistic target of rapamycin (mTOR) kinase forms two multi-protein signaling complexes, mTORC1 and mTORC2, which are master regulators of cell growth, metabolism, survival and autophagy. Two of the subunits of these complexes are mLST8 and Raptor, β-propeller proteins that stabilize the mTOR...

Descripción completa

Detalles Bibliográficos
Autores principales: Cuéllar, Jorge, Ludlam, W. Grant, Tensmeyer, Nicole C., Aoba, Takuma, Dhavale, Madhura, Santiago, César, Bueno-Carrasco, M. Teresa, Mann, Michael J., Plimpton, Rebecca L., Makaju, Aman, Franklin, Sarah, Willardson, Barry M., Valpuesta, José M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6599039/
https://www.ncbi.nlm.nih.gov/pubmed/31253771
http://dx.doi.org/10.1038/s41467-019-10781-1
_version_ 1783430879134089216
author Cuéllar, Jorge
Ludlam, W. Grant
Tensmeyer, Nicole C.
Aoba, Takuma
Dhavale, Madhura
Santiago, César
Bueno-Carrasco, M. Teresa
Mann, Michael J.
Plimpton, Rebecca L.
Makaju, Aman
Franklin, Sarah
Willardson, Barry M.
Valpuesta, José M.
author_facet Cuéllar, Jorge
Ludlam, W. Grant
Tensmeyer, Nicole C.
Aoba, Takuma
Dhavale, Madhura
Santiago, César
Bueno-Carrasco, M. Teresa
Mann, Michael J.
Plimpton, Rebecca L.
Makaju, Aman
Franklin, Sarah
Willardson, Barry M.
Valpuesta, José M.
author_sort Cuéllar, Jorge
collection PubMed
description The mechanistic target of rapamycin (mTOR) kinase forms two multi-protein signaling complexes, mTORC1 and mTORC2, which are master regulators of cell growth, metabolism, survival and autophagy. Two of the subunits of these complexes are mLST8 and Raptor, β-propeller proteins that stabilize the mTOR kinase and recruit substrates, respectively. Here we report that the eukaryotic chaperonin CCT plays a key role in mTORC assembly and signaling by folding both mLST8 and Raptor. A high resolution (4.0 Å) cryo-EM structure of the human mLST8-CCT intermediate isolated directly from cells shows mLST8 in a near-native state bound to CCT deep within the folding chamber between the two CCT rings, and interacting mainly with the disordered N- and C-termini of specific CCT subunits of both rings. These findings describe a unique function of CCT in mTORC assembly and a distinct binding site in CCT for mLST8, far from those found for similar β-propeller proteins.
format Online
Article
Text
id pubmed-6599039
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-65990392019-07-01 Structural and functional analysis of the role of the chaperonin CCT in mTOR complex assembly Cuéllar, Jorge Ludlam, W. Grant Tensmeyer, Nicole C. Aoba, Takuma Dhavale, Madhura Santiago, César Bueno-Carrasco, M. Teresa Mann, Michael J. Plimpton, Rebecca L. Makaju, Aman Franklin, Sarah Willardson, Barry M. Valpuesta, José M. Nat Commun Article The mechanistic target of rapamycin (mTOR) kinase forms two multi-protein signaling complexes, mTORC1 and mTORC2, which are master regulators of cell growth, metabolism, survival and autophagy. Two of the subunits of these complexes are mLST8 and Raptor, β-propeller proteins that stabilize the mTOR kinase and recruit substrates, respectively. Here we report that the eukaryotic chaperonin CCT plays a key role in mTORC assembly and signaling by folding both mLST8 and Raptor. A high resolution (4.0 Å) cryo-EM structure of the human mLST8-CCT intermediate isolated directly from cells shows mLST8 in a near-native state bound to CCT deep within the folding chamber between the two CCT rings, and interacting mainly with the disordered N- and C-termini of specific CCT subunits of both rings. These findings describe a unique function of CCT in mTORC assembly and a distinct binding site in CCT for mLST8, far from those found for similar β-propeller proteins. Nature Publishing Group UK 2019-06-28 /pmc/articles/PMC6599039/ /pubmed/31253771 http://dx.doi.org/10.1038/s41467-019-10781-1 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Cuéllar, Jorge
Ludlam, W. Grant
Tensmeyer, Nicole C.
Aoba, Takuma
Dhavale, Madhura
Santiago, César
Bueno-Carrasco, M. Teresa
Mann, Michael J.
Plimpton, Rebecca L.
Makaju, Aman
Franklin, Sarah
Willardson, Barry M.
Valpuesta, José M.
Structural and functional analysis of the role of the chaperonin CCT in mTOR complex assembly
title Structural and functional analysis of the role of the chaperonin CCT in mTOR complex assembly
title_full Structural and functional analysis of the role of the chaperonin CCT in mTOR complex assembly
title_fullStr Structural and functional analysis of the role of the chaperonin CCT in mTOR complex assembly
title_full_unstemmed Structural and functional analysis of the role of the chaperonin CCT in mTOR complex assembly
title_short Structural and functional analysis of the role of the chaperonin CCT in mTOR complex assembly
title_sort structural and functional analysis of the role of the chaperonin cct in mtor complex assembly
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6599039/
https://www.ncbi.nlm.nih.gov/pubmed/31253771
http://dx.doi.org/10.1038/s41467-019-10781-1
work_keys_str_mv AT cuellarjorge structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT ludlamwgrant structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT tensmeyernicolec structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT aobatakuma structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT dhavalemadhura structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT santiagocesar structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT buenocarrascomteresa structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT mannmichaelj structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT plimptonrebeccal structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT makajuaman structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT franklinsarah structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT willardsonbarrym structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly
AT valpuestajosem structuralandfunctionalanalysisoftheroleofthechaperonincctinmtorcomplexassembly