PMID- 12370290
OWN - NLM
STAT- MEDLINE
DCOM- 20021122
LR  - 20211203
IS  - 0270-7306 (Print)
IS  - 1098-5549 (Electronic)
IS  - 0270-7306 (Linking)
VI  - 22
IP  - 21
DP  - 2002 Nov
TI  - The rapamycin-binding domain governs substrate selectivity by the mammalian 
      target of rapamycin.
PG  - 7428-38
AB  - The mammalian target of rapamycin (mTOR) is a Ser/Thr (S/T) protein kinase, which 
      controls mRNA translation initiation by modulating phosphorylation of the 
      translational regulators PHAS-I and p70(S6K). Here we show that in vitro mTOR is 
      able to phosphorylate these two regulators at comparable rates. Both (S/T)P 
      sites, such as Thr36, Thr45, and Thr69 in PHAS-I and the h(S/T)h site (where h is 
      a hydrophobic amino acid) Thr389 in p70(S6K), were phosphorylated. 
      Rapamycin-FKBP12 inhibited mTOR activity. Surprisingly, the extent of inhibition 
      depended on the substrate. Moreover, mutating Ser2035 in the rapamycin-binding 
      domain (FRB) not only decreased rapamycin sensitivity as expected but also 
      dramatically affected the sites phosphorylated by mTOR. The results demonstrate 
      that mutations in Ser2035 are not silent with respect to mTOR activity and 
      implicate the FRB in substrate recognition. The findings also impose new 
      limitations on interpreting results from experiments in which rapamycin and/or 
      rapamycin-resistant forms of mTOR are used to investigate mTOR function in cells.
FAU - McMahon, Lloyd P
AU  - McMahon LP
AD  - Department of Pharmacology, University of Virginia School of Medicine, 
      Charlottesville, Virginia 22908, USA.
FAU - Choi, Kin M
AU  - Choi KM
FAU - Lin, Tai-An
AU  - Lin TA
FAU - Abraham, Robert T
AU  - Abraham RT
FAU - Lawrence, John C Jr
AU  - Lawrence JC Jr
LA  - eng
GR  - DK52753/DK/NIDDK NIH HHS/United States
GR  - CA 76193/CA/NCI NIH HHS/United States
GR  - R01 DK028312/DK/NIDDK NIH HHS/United States
GR  - DK28312/DK/NIDDK NIH HHS/United States
GR  - R01 DK052753/DK/NIDDK NIH HHS/United States
GR  - R01 CA076193/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Mol Cell Biol
JT  - Molecular and cellular biology
JID - 8109087
RN  - 0 (Adaptor Proteins, Signal Transducing)
RN  - 0 (Antibiotics, Antineoplastic)
RN  - 0 (Carrier Proteins)
RN  - 0 (Cell Cycle Proteins)
RN  - 0 (DNA, Complementary)
RN  - 0 (EIF4EBP1 protein, human)
RN  - 0 (Phosphoproteins)
RN  - 0 (Recombinant Proteins)
RN  - 2ZD004190S (Threonine)
RN  - 452VLY9402 (Serine)
RN  - EC 2.7.- (Protein Kinases)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.11.1 (Ribosomal Protein S6 Kinases, 70-kDa)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - W36ZG6FT64 (Sirolimus)
SB  - IM
MH  - Adaptor Proteins, Signal Transducing
MH  - Antibiotics, Antineoplastic/*pharmacology
MH  - Binding Sites
MH  - Carrier Proteins/metabolism
MH  - Cell Cycle Proteins
MH  - Cell Line
MH  - DNA, Complementary/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Humans
MH  - Kinetics
MH  - Models, Biological
MH  - Mutagenesis, Site-Directed
MH  - Mutation
MH  - Phosphoproteins/metabolism
MH  - Phosphorylation
MH  - Protein Binding
MH  - Protein Kinases/metabolism/*physiology
MH  - Protein Structure, Tertiary
MH  - Recombinant Proteins/metabolism
MH  - Ribosomal Protein S6 Kinases, 70-kDa/metabolism
MH  - Serine/chemistry/metabolism
MH  - Sirolimus/*pharmacology
MH  - Substrate Specificity
MH  - TOR Serine-Threonine Kinases
MH  - Threonine/chemistry
MH  - Time Factors
MH  - Transfection
PMC - PMC135667
EDAT- 2002/10/09 04:00
MHDA- 2002/11/26 04:00
PMCR- 2002/11/01
CRDT- 2002/10/09 04:00
PHST- 2002/10/09 04:00 [pubmed]
PHST- 2002/11/26 04:00 [medline]
PHST- 2002/10/09 04:00 [entrez]
PHST- 2002/11/01 00:00 [pmc-release]
AID - 0562 [pii]
AID - 10.1128/MCB.22.21.7428-7438.2002 [doi]
PST - ppublish
SO  - Mol Cell Biol. 2002 Nov;22(21):7428-38. doi: 10.1128/MCB.22.21.7428-7438.2002.