PMID- 24337580
OWN - NLM
STAT- MEDLINE
DCOM- 20140410
LR  - 20211203
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 289
IP  - 5
DP  - 2014 Jan 31
TI  - Amino acids activate mammalian target of rapamycin (mTOR) complex 1 without 
      changing Rag GTPase guanyl nucleotide charging.
PG  - 2658-74
LID - 10.1074/jbc.M113.528505 [doi]
AB  - Activation of mammalian target of rapamycin complex 1 (mTORC1) by amino acids is 
      mediated in part by the Rag GTPases, which bind the raptor subunit of mTORC1 in 
      an amino acid-stimulated manner and promote mTORC1 interaction with Rheb-GTP, the 
      immediate activator. Here we examine whether the ability of amino acids to 
      regulate mTORC1 binding to Rag and mTORC1 activation is due to the regulation of 
      Rag guanyl nucleotide charging. Rag heterodimers in vitro exhibit a very rapid, 
      spontaneous exchange of guanyl nucleotides and an inability to hydrolyze GTP. 
      Mutation of the Rag P-loop corresponding to Ras(Ser-17) abolishes guanyl 
      nucleotide binding. Such a mutation in RagA or RagB inhibits, whereas in RagC or 
      RagD it enhances, Rag heterodimer binding to mTORC1. The binding of wild-type and 
      mutant Rag heterodimers to mTORC1 in vitro parallels that seen with transient 
      expression, but binding to mTORC1 in vitro is entirely independent of Rag guanyl 
      nucleotide charging. HeLa cells stably overexpressing wild-type or P-loop mutant 
      RagC exhibit unaltered amino acid regulation of mTORC1. Despite amino 
      acid-independent raptor binding to Rag, mTORC1 is inhibited by amino acid 
      withdrawal as in parental cells. Rag heterodimers extracted from (32)P-labeled 
      whole cells, or just from the pool associated with the lysosomal membrane, 
      exhibit constitutive [(32)P]GTP charging that is unaltered by amino acid 
      withdrawal. Thus, amino acids promote mTORC1 activation without altering Rag GTP 
      charging. Raptor binding to Rag, although necessary, is not sufficient for mTORC1 
      activation. Additional amino acid-dependent steps couple Rag-mTORC1 to Rheb-GTP.
FAU - Oshiro, Noriko
AU  - Oshiro N
AD  - From the Department of Molecular Biology and Diabetes Unit, Medical Services, 
      Massachusetts General Hospital, Boston, Massachusetts 02114 and the Department of 
      Medicine, Harvard Medical School, Boston, Massachusetts 02115.
FAU - Rapley, Joseph
AU  - Rapley J
FAU - Avruch, Joseph
AU  - Avruch J
LA  - eng
GR  - P30 DK040561/DK/NIDDK NIH HHS/United States
GR  - P30DK057521/DK/NIDDK NIH HHS/United States
GR  - R01CA73818/CA/NCI NIH HHS/United States
GR  - P30 DK057521/DK/NIDDK NIH HHS/United States
GR  - R37DK17776/DK/NIDDK NIH HHS/United States
GR  - R01 CA073818/CA/NCI NIH HHS/United States
GR  - R37 DK017776/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20131211
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Adaptor Proteins, Signal Transducing)
RN  - 0 (Amino Acids)
RN  - 0 (MLST8 protein, human)
RN  - 0 (Multiprotein Complexes)
RN  - 0 (RPTOR protein, human)
RN  - 0 (RRAGC protein, human)
RN  - 0 (RRAGD protein, human)
RN  - 0 (Regulatory-Associated Protein of mTOR)
RN  - 0 (mTOR Associated Protein, LST8 Homolog)
RN  - 10028-17-8 (Tritium)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.11.1 (Mechanistic Target of Rapamycin Complex 1)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - EC 3.6.1.- (GTP Phosphohydrolases)
RN  - EC 3.6.1.- (RRAGA protein, human)
RN  - EC 3.6.1.- (RRAGB protein, human)
RN  - EC 3.6.5.2 (Monomeric GTP-Binding Proteins)
SB  - IM
MH  - Adaptor Proteins, Signal Transducing/metabolism
MH  - Amino Acids/*metabolism
MH  - Dimerization
MH  - Enzyme Activation/physiology
MH  - GTP Phosphohydrolases/chemistry/genetics/*metabolism
MH  - HEK293 Cells
MH  - HeLa Cells
MH  - Humans
MH  - Mechanistic Target of Rapamycin Complex 1
MH  - Monomeric GTP-Binding Proteins/chemistry/genetics/*metabolism
MH  - Multiprotein Complexes/*metabolism
MH  - Protein Binding/physiology
MH  - Regulatory-Associated Protein of mTOR
MH  - Signal Transduction/physiology
MH  - TOR Serine-Threonine Kinases/*metabolism
MH  - Tritium
MH  - mTOR Associated Protein, LST8 Homolog
PMC - PMC3908400
OTO - NOTNLM
OT  - Amino Acids
OT  - Cell Signaling
OT  - GTPase
OT  - Insulin
OT  - Lysosomes
OT  - Rag GTPase
OT  - Raptor
OT  - mTOR Complex (mTORC)
EDAT- 2013/12/18 06:00
MHDA- 2014/04/11 06:00
PMCR- 2013/12/11
CRDT- 2013/12/17 06:00
PHST- 2013/12/17 06:00 [entrez]
PHST- 2013/12/18 06:00 [pubmed]
PHST- 2014/04/11 06:00 [medline]
PHST- 2013/12/11 00:00 [pmc-release]
AID - S0021-9258(19)74788-7 [pii]
AID - M113.528505 [pii]
AID - 10.1074/jbc.M113.528505 [doi]
PST - ppublish
SO  - J Biol Chem. 2014 Jan 31;289(5):2658-74. doi: 10.1074/jbc.M113.528505. Epub 2013 
      Dec 11.