PMID- 31253771
OWN - NLM
STAT- MEDLINE
DCOM- 20190827
LR  - 20211204
IS  - 2041-1723 (Electronic)
IS  - 2041-1723 (Linking)
VI  - 10
IP  - 1
DP  - 2019 Jun 28
TI  - Structural and functional analysis of the role of the chaperonin CCT in mTOR 
      complex assembly.
PG  - 2865
LID - 10.1038/s41467-019-10781-1 [doi]
LID - 2865
AB  - 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, beta-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 A) 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 
      beta-propeller proteins.
FAU - Cuellar, Jorge
AU  - Cuellar J
AD  - Centro Nacional de Biotecnologia, Campus de la Universidad Autonoma de Madrid, 
      28049, Madrid, Spain.
FAU - Ludlam, W Grant
AU  - Ludlam WG
AUID- ORCID: 0000-0002-2701-8791
AD  - Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 
      84602, USA.
FAU - Tensmeyer, Nicole C
AU  - Tensmeyer NC
AUID- ORCID: 0000-0003-4957-7129
AD  - Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 
      84602, USA.
FAU - Aoba, Takuma
AU  - Aoba T
AD  - Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 
      84602, USA.
FAU - Dhavale, Madhura
AU  - Dhavale M
AD  - Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 
      84602, USA.
FAU - Santiago, Cesar
AU  - Santiago C
AD  - Centro Nacional de Biotecnologia, Campus de la Universidad Autonoma de Madrid, 
      28049, Madrid, Spain.
FAU - Bueno-Carrasco, M Teresa
AU  - Bueno-Carrasco MT
AD  - Centro Nacional de Biotecnologia, Campus de la Universidad Autonoma de Madrid, 
      28049, Madrid, Spain.
FAU - Mann, Michael J
AU  - Mann MJ
AD  - Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 
      84602, USA.
FAU - Plimpton, Rebecca L
AU  - Plimpton RL
AD  - Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 
      84602, USA.
FAU - Makaju, Aman
AU  - Makaju A
AD  - Department of Internal Medicine, Nora Eccles Harrison Cardiovascular Research and 
      Training Institute, University of Utah, Salt Lake City, UT, 84112, USA.
FAU - Franklin, Sarah
AU  - Franklin S
AD  - Department of Internal Medicine, Nora Eccles Harrison Cardiovascular Research and 
      Training Institute, University of Utah, Salt Lake City, UT, 84112, USA.
FAU - Willardson, Barry M
AU  - Willardson BM
AD  - Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 
      84602, USA. bmwillardson@chem.byu.edu.
FAU - Valpuesta, Jose M
AU  - Valpuesta JM
AUID- ORCID: 0000-0001-7468-8053
AD  - Centro Nacional de Biotecnologia, Campus de la Universidad Autonoma de Madrid, 
      28049, Madrid, Spain. jmv@cnb.csic.es.
LA  - eng
GR  - R01 EY012287/EY/NEI NIH HHS/United States
GR  - EY012287/U.S. Department of Health & Human Services | National Institutes of 
      Health (NIH)/International
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20190628
PL  - England
TA  - Nat Commun
JT  - Nature communications
JID - 101528555
RN  - 0 (MLST8 protein, human)
RN  - 0 (RPTOR protein, human)
RN  - 0 (Regulatory-Associated Protein of mTOR)
RN  - 0 (mTOR Associated Protein, LST8 Homolog)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - EC 3.6.1.- (Chaperonin Containing TCP-1)
SB  - IM
MH  - Amino Acid Sequence
MH  - Binding Sites
MH  - Chaperonin Containing TCP-1/*physiology
MH  - Cryoelectron Microscopy
MH  - Gene Expression Regulation/physiology
MH  - Gene Knockdown Techniques
MH  - HEK293 Cells
MH  - Hep G2 Cells
MH  - Humans
MH  - Mass Spectrometry
MH  - Models, Molecular
MH  - Protein Binding
MH  - Protein Conformation
MH  - Protein Folding
MH  - Regulatory-Associated Protein of mTOR/genetics/*metabolism
MH  - TOR Serine-Threonine Kinases/genetics/*metabolism
MH  - mTOR Associated Protein, LST8 Homolog/genetics/*metabolism
PMC - PMC6599039
COIS- The authors declare no competing interests.
EDAT- 2019/06/30 06:00
MHDA- 2019/08/28 06:00
PMCR- 2019/06/28
CRDT- 2019/06/30 06:00
PHST- 2019/02/26 00:00 [received]
PHST- 2019/06/01 00:00 [accepted]
PHST- 2019/06/30 06:00 [entrez]
PHST- 2019/06/30 06:00 [pubmed]
PHST- 2019/08/28 06:00 [medline]
PHST- 2019/06/28 00:00 [pmc-release]
AID - 10.1038/s41467-019-10781-1 [pii]
AID - 10781 [pii]
AID - 10.1038/s41467-019-10781-1 [doi]
PST - epublish
SO  - Nat Commun. 2019 Jun 28;10(1):2865. doi: 10.1038/s41467-019-10781-1.