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Section I: Research Areas<br />
chapter 02: Signaling<br />
mTOR Signaling<br />
GRB10<br />
Sin1<br />
Torin1<br />
PP242<br />
KU63794<br />
WYE354<br />
PRR5<br />
mTORC2<br />
Rictor GβL<br />
mTOR<br />
DEPTOR<br />
SGK1<br />
rapamycin<br />
FKBP12<br />
PKCα<br />
PRAS40<br />
p70S6K<br />
Cell Growth<br />
IRS-1<br />
Akt<br />
TSC1<br />
TSC2<br />
TBC1D7<br />
Rheb<br />
mTORC1<br />
GβL<br />
Raptor<br />
mTOR<br />
DEPTOR<br />
FIP200<br />
Atg13<br />
ULK<br />
Glucose<br />
PIP 3 PIP 2<br />
AMP: ATP<br />
Growth Factors,<br />
Hormones,<br />
AICAR<br />
Cytokines, etc.<br />
Stress<br />
Hypoxia<br />
PI3K<br />
Dvl<br />
DNA<br />
Damage<br />
PDK1<br />
Ras<br />
PTEN<br />
p53<br />
Autophagy<br />
eIF4G<br />
GATOR2<br />
54 For Research Use Only. Not For Use in Diagnostic Procedures. See pages 302 & 303 for Pathway Diagrams, Application, and Reactivity keys.<br />
4E-<br />
BP1/2<br />
mRNA<br />
Translation<br />
Proliferation<br />
LRP<br />
Erk<br />
RSK<br />
GSK-3<br />
REDD1/2<br />
AMPK<br />
Lipin 1<br />
Wnt<br />
Frizzled<br />
Gα q/o<br />
mTORC1<br />
Translocation<br />
to Lysosome<br />
Lipid Synthesis<br />
LKB1<br />
Sestrin-1/2<br />
LAMTOR<br />
1/2/3/4/5<br />
Ragulator<br />
Complex<br />
Rag A/B<br />
GTP<br />
Rag C/D<br />
GDP<br />
Ribosome<br />
Biogenesis<br />
TFEB<br />
PPARα<br />
HIF-1<br />
PGC-1α<br />
PPARγ<br />
SKAR<br />
mRNA Splicing<br />
metformin<br />
Glucose,<br />
Amino Acids<br />
V-ATPase<br />
Mios<br />
WDR59<br />
Lipogenesis<br />
SREBP-1<br />
Transcription<br />
Seh1L<br />
WDR24<br />
Sec13<br />
GATOR1<br />
DEPDC5 Nprl2<br />
Nprl3<br />
Lipid<br />
Metabolism<br />
FLCN<br />
FNIP1/2<br />
Autophagy/Lysosome<br />
Biogenesis<br />
VEGF/<br />
Angiogenesis<br />
Mitochondrial<br />
Metabolism<br />
Adipogenesis<br />
The mechanistic target of rapamycin (mTOR) is an atypical serine/threonine kinase that is present in two distinct complexes. The first, mTOR complex 1 (mTORC1), is<br />
composed of mTOR, Raptor, GβL, and DEPTOR and is inhibited by rapamycin. It is a master growth regulator that senses and integrates diverse nutritional and environmental<br />
cues, including growth factors, energy levels, cellular stress, and amino acids. It couples these signals to the promotion of cellular growth by phosphorylating substrates<br />
that potentiate anabolic processes such as mRNA translation and lipid synthesis, or limit catabolic processes such as autophagy. The small GTPase Rheb, in its GTP-bound<br />
state, is a necessary and potent stimulator of mTORC1 kinase activity, which is negatively regulated by its GAP, the tuberous sclerosis heterodimer TSC1/2. Most upstream<br />
inputs are funneled through Akt and TSC1/2 to regulate the nucleotide-loading state of Rheb. In contrast, amino acids signal to mTORC1 independently of the PI3K/Akt axis to<br />
promote the translocation of mTORC1 to the lysosomal surface where it can become activated upon contact with Rheb. This process is mediated by the coordinated actions of<br />
multiple complexes, notably the v-ATPase, Ragulator, the Rag GTPases, and GATOR1/2. The second complex, mTOR complex 2 (mTORC2), is composed of mTOR, Rictor, GβL,<br />
Sin1, PRR5/Protor-1, and DEPTOR. mTORC2 promotes cellular survival by activating Akt, regulates cytoskeletal dynamics by activating PKCα, and controls ion transport and<br />
growth via SGK1 phosphorylation. Aberrant mTOR signaling is involved in many disease states including cancer, cardiovascular disease, and diabetes.<br />
Select Reviews:<br />
Dowling, R.J., Topisirovic, I., Fonseca, B.D., and Sonenberg, N. (2010) Biochim. Biophys. Acta. 1804, 433–439. • Dunlop, E.A. and Tee, A.R. (2009) Cell. Signal. 21,<br />
827–835. • Hoeffer, C.A. and Klann, E. (2010) Trends Neurosci. 33, 67–75. • Laplante, M. and Sabatini, D.M. (2013) J. Cell Sci. 126, 1713–1719. • Laplante, M. and<br />
Sabatini, D.M. (2012) Cell 149, 274–293. • Neufeld, T.P. (2010) Curr. Opin. Cell Biol. 22, 157–168. • Zoncu, R., Efeyan, A., and Sabatini, D.M. (2011) Nat. Rev. Mol. Cell<br />
Biol. 12, 21–35.<br />
© 2002–2015 Cell Signaling Technology, Inc. • We would like to thank Rachel Wolfson and Prof. David Sabatini, Whitehead Institute for Biomedical Research, MIT, Cambridge, MA, for reviewing this diagram.<br />
Akt Substrates<br />
MAPK, mTOR, and the PI3K/Akt pathways are key signaling pathways activated downstream of oncogenic receptor tyrosine<br />
kinases (RTKs). All of these pathways activate AGC kinase family members, including Akt, RSK, and p70 S6 kinases, whose<br />
protein substrates are phosphorylated at the RxRxxS/T motif.<br />
In a phosphoproteomic study co-authored by scientists in the Cell Signaling Technology (<strong>CST</strong>) Site Discovery Group (Moritz, A.<br />
et al. (2010) Sci. Signal 24,ra64), over 300 novel downstream substrates for these AGC family kinases were identified. The<br />
experimental approach involved the use of PhosphoScan ® , <strong>CST</strong>’s proprietary methodology for antibody-based peptide enrichment<br />
combined with tandem mass spectrometry for quantitative profiling of post-translational modifications. A key step was<br />
the development of a RxRxxS/T motif antibody, which was then used as an affinity reagent to selectively immunoprecipitate<br />
phosphorylated substrates of Akt, RSK, and p70 S6 kinases. The antibody was employed in PhosphoScan in three different<br />
cancer cell lines, dependent on either EGFR, c-Met, or PDGFR, allowing mapping of the signaling network downstream of these<br />
RTKs. Substrates included proteins involved in many cellular functions, including scaffolding, protein stability, metabolism, trafficking,<br />
and motility.<br />
Substrate<br />
GAP/GEF/Adaptors<br />
ARHGAP19<br />
ARHGEF12<br />
AS250<br />
TBC1D1<br />
TBC1D4<br />
Receptors/Transporters<br />
DR6 SLC20A2<br />
EPHA2 SLC9A1<br />
FGFR2<br />
SEMA4B<br />
TSC2<br />
CD2AP<br />
FRS2<br />
IRS1<br />
IRS2<br />
Adhesion/Cytoskeleton<br />
DSP PLEC1<br />
MLLT4 SVIL<br />
KIF21A PPP1R12A<br />
AMPKA<br />
Isoform<br />
RICTOR<br />
CABLES1<br />
LMO7<br />
Kinases<br />
WNK1<br />
PKD2<br />
HGK<br />
BRD2<br />
UO126<br />
EGFR<br />
Gefitinib Su11274 Gleevec<br />
Ras<br />
MEK<br />
RSK<br />
Energy/<br />
Metabolism<br />
PANK2<br />
PFKFB2<br />
OXR1<br />
GSK-3α<br />
GSK-3β<br />
PI3K<br />
Akt<br />
Met<br />
SGKs<br />
mTOR<br />
Wort<br />
S6K<br />
PDGFRα<br />
RNA Processing/<br />
Translation<br />
LARP1<br />
MEPCE<br />
RPS6<br />
EIF4ENIF1<br />
EDC3<br />
Published Data Human<br />
Organism Site Site Sequence (+/-7) PMID<br />
Rapa<br />
Chaperone/Ubiquitin<br />
CCT2<br />
DNAJC2<br />
SGTA<br />
NEDD4-2<br />
UBR4<br />
UBXN4<br />
NIPA<br />
TIF1-γ<br />
Survival<br />
AKT1S1<br />
BAD<br />
NDRG2<br />
NDRG3<br />
Transcription<br />
FOXO3<br />
GTF3C1<br />
IWS1<br />
TAF3<br />
TCF12<br />
Vesicle Trafficking<br />
C4orf16 REPS1<br />
GOLGA4 STX12<br />
NDRG1 STX7<br />
HDGF2<br />
TCF3<br />
BRD1<br />
SP100<br />
Substrate Function and<br />
Effect of Phosphorylation<br />
14-3-3 z Akt1 human S58 S58 VVGARRSsWRVVssI 11956222 A key regulatory protein in signal transduction,<br />
checkpoint control, apoptotic,<br />
and nutrient-sensing pathways; effect<br />
of phosphorylation is unknown<br />
acinus Akt1 human S1180 S1180 GPRsRsRsRDRRRKE 18559500,<br />
16177823<br />
Akt1 rat S1329 S1331 HSRSRSRsTPVRDRG 16177823<br />
Induces chromatin condensation during<br />
apoptosis; phosphorylation inhibits<br />
this process<br />
ACLY Akt1 mouse S455 S455 PAPSRtAsFsESRAD 16007201 Catalyzes the formation of acetyl-CoA<br />
and oxaloacetate (OAA) in the cytosol;<br />
phosphorylation enhances the catalytic<br />
activity of the enzyme<br />
ADRB2 Akt1 human S346 S346 LLCLRRssLKAyGNG 11809767 A receptor that binds epinephrine<br />
and norepinephrine, acting as a<br />
neuromodulator in the central nervous<br />
system and as a hormone in the<br />
vascular system; phosphorylation in<br />
response to insulin stimulation leads to<br />
sequestration of ADBR2<br />
Akt1 Akt1 human S246,<br />
T72<br />
S246,<br />
T72<br />
LSRERVFsEDRARFY,<br />
TERPRPNtFIIRCLQ<br />
Akt1 mouse S473 S473 RPHFPQFsYsAsGtA 11570877,<br />
10722653<br />
16549426 Activated by insulin and various growth<br />
and survival factors to function in a<br />
wortmannin-sensitive PI3 kinaseinvolved<br />
pathway controlling survival<br />
and apoptosis; autophosphorylation<br />
activates the kinase<br />
AMPKA1 Akt1 rat S485 S485 ATPQRSGsISNYRSC 16340011 Heterotrimeric complex that plays a<br />
key role in the regulation of energy<br />
homeostasis; phosphorylation regulates<br />
AMPK activity<br />
AMPKA2 Akt1 rat S491 S491 STPQRSCsAAGLHRP 16340011 Heterotrimeric complex that plays a<br />
key role in the regulation of energy<br />
homeostasis; phosphorylation regulates<br />
AMPK activity<br />
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