PMID- 20022946
OWN - NLM
STAT- MEDLINE
DCOM- 20100415
LR  - 20211203
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 285
IP  - 11
DP  - 2010 Mar 12
TI  - mTOR Ser-2481 autophosphorylation monitors mTORC-specific catalytic activity and 
      clarifies rapamycin mechanism of action.
PG  - 7866-79
LID - 10.1074/jbc.M109.096222 [doi]
AB  - The mammalian target of rapamycin (mTOR) Ser/Thr kinase signals in at least two 
      multiprotein complexes distinguished by their different partners and 
      sensitivities to rapamycin. Acute rapamycin inhibits signaling by mTOR complex 1 
      (mTORC1) but not mTOR complex 2 (mTORC2), which both promote cell growth, 
      proliferation, and survival. Although mTORC2 regulation remains poorly defined, 
      diverse cellular mitogens activate mTORC1 signaling in a manner that requires 
      sufficient levels of amino acids and cellular energy. Before the identification 
      of distinct mTOR complexes, mTOR was reported to autophosphorylate on Ser-2481 in 
      vivo in a rapamycin- and amino acid-insensitive manner. These results suggested 
      that modulation of mTOR intrinsic catalytic activity does not universally 
      underlie mTOR regulation. Here we re-examine the regulation of mTOR Ser-2481 
      autophosphorylation (Ser(P)-2481) in vivo by studying mTORC-specific Ser(P)-2481 
      in mTORC1 and mTORC2, with a primary focus on mTORC1. In contrast to previous 
      work, we find that acute rapamycin and amino acid withdrawal markedly attenuate 
      mTORC1-associated mTOR Ser(P)-2481 in cycling cells. Although insulin stimulates 
      both mTORC1- and mTORC2-associated mTOR Ser(P)-2481 in a phosphatidylinositol 
      3-kinase-dependent manner, rapamycin acutely inhibits insulin-stimulated mTOR 
      Ser(P)-2481 in mTORC1 but not mTORC2. By interrogating diverse mTORC1 regulatory 
      input, we find that without exception mTORC1-activating signals promote, whereas 
      mTORC1-inhibitory signals decrease mTORC1-associated mTOR Ser(P)-2481. These data 
      suggest that mTORC1- and likely mTORC2-associated mTOR Ser-2481 
      autophosphorylation directly monitors intrinsic mTORC-specific catalytic activity 
      and reveal that rapamycin inhibits mTORC1 signaling in vivo by reducing mTORC1 
      catalytic activity.
FAU - Soliman, Ghada A
AU  - Soliman GA
AD  - Department of Cell and Developmental Biology, University of Michigan Medical 
      School, Ann Arbor, Michigan 48109-2200, USA.
FAU - Acosta-Jaquez, Hugo A
AU  - Acosta-Jaquez HA
FAU - Dunlop, Elaine A
AU  - Dunlop EA
FAU - Ekim, Bilgen
AU  - Ekim B
FAU - Maj, Nicole E
AU  - Maj NE
FAU - Tee, Andrew R
AU  - Tee AR
FAU - Fingar, Diane C
AU  - Fingar DC
LA  - eng
GR  - K01 DK060654/DK/NIDDK NIH HHS/United States
GR  - R01 DK078135/DK/NIDDK NIH HHS/United States
GR  - R01 DK-078135/DK/NIDDK NIH HHS/United States
GR  - 06-0914/AICR_/Worldwide Cancer Research/United Kingdom
GR  - K01 DK060654-03/DK/NIDDK NIH HHS/United States
GR  - P60 DK020572/DK/NIDDK NIH HHS/United States
GR  - K01 DK-60654/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20091218
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Antibiotics, Antineoplastic)
RN  - 0 (Antibodies)
RN  - 0 (CRTC2 protein, human)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Multiprotein Complexes)
RN  - 0 (Proteins)
RN  - 0 (Transcription Factors)
RN  - 452VLY9402 (Serine)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.1.1 (mTOR protein, mouse)
RN  - EC 2.7.11.1 (Mechanistic Target of Rapamycin Complex 1)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - W36ZG6FT64 (Sirolimus)
SB  - IM
MH  - 3T3-L1 Cells
MH  - Animals
MH  - Antibiotics, Antineoplastic/*pharmacology
MH  - Antibodies/pharmacology
MH  - Catalysis
MH  - Cell Line, Transformed
MH  - Fibroblasts/cytology
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins/immunology/*metabolism
MH  - Kidney/cytology
MH  - Mechanistic Target of Rapamycin Complex 1
MH  - Mice
MH  - Multiprotein Complexes
MH  - Phosphorylation
MH  - Protein Serine-Threonine Kinases/immunology/*metabolism
MH  - Proteins
MH  - Rabbits
MH  - Serine/metabolism
MH  - Signal Transduction/*physiology
MH  - Sirolimus/*pharmacology
MH  - TOR Serine-Threonine Kinases
MH  - Transcription Factors/metabolism
PMC - PMC2832937
EDAT- 2009/12/22 06:00
MHDA- 2010/04/16 06:00
PMCR- 2011/03/12
CRDT- 2009/12/22 06:00
PHST- 2009/12/22 06:00 [entrez]
PHST- 2009/12/22 06:00 [pubmed]
PHST- 2010/04/16 06:00 [medline]
PHST- 2011/03/12 00:00 [pmc-release]
AID - S0021-9258(19)64425-X [pii]
AID - M109.096222 [pii]
AID - 10.1074/jbc.M109.096222 [doi]
PST - ppublish
SO  - J Biol Chem. 2010 Mar 12;285(11):7866-79. doi: 10.1074/jbc.M109.096222. Epub 2009 
      Dec 18.