The principal target of rapamycin‐induced p70s6k inactivation is a novel phosphorylation site within a conserved hydrophobic domain.
Open accessAbstract
The immunosuppressive agent rapamycin induces inactivation of p70s6k with no effect on other mitogen-activated kinases. Here we have employed a combination of techniques, including mass spectrometry, to demonstrate that this effect is associated with selective dephosphorylation of three previously unidentified p70s6k phosphorylation sites: T229, T389 and S404. T229 resides at a conserved position in the catalytic domain, whose phosphorylation is essential for the activation of other mitogen-induced kinases. However, the principal target of rapamycin-induced p70s6k inactivation is T389, which is located in an unusual hydrophobic sequence outside the catalytic domain. Mutation of T389 to alanine ablates kinase activity, whereas mutation to glutamic acid confers constitutive kinase activity and rapamycin resistance. The importance of this site and its surrounding motif to kinase function is emphasized by its presence in a large number of protein kinases of the second messenger family and its conservation in putative p70s6k homologues from as distantly related organisms as yeast and plants.
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mTOR phosphorylates and activates p70S6K at threonine389 and is a marker of mTOR activity.
“mTOR phosphorylates and activates p70S6K at threonine389 and is a marker of mTOR activity [40].”
From Tuberous Sclerosis Protein 2 (TSC2) Modulates CCN4 Cytoprotection During Apoptotic Amyloid Toxicity in Microglia (2013) · cites this paper as [40]
The mutational-analysis section finds that phosphorylation of p70S6K at T389 is critical for its activity. The Discussion does not establish that mTOR (FRAP/RAFT1) directly phosphorylates that site: it says FRAP/RAFT1's upstream role remained to be determined and considers a phosphatase-based alternative. The paper also does not identify T389 phosphorylation as a marker of mTOR activity. Page numbers were not available in our copy. (LLM, conf=0.88)
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Research works printed but not linked: 14
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BackgroundInfluential
- Authors
- Pearson, R. B., Dennis, P. B., Han, J. W., Williamson, N. A., Kozma, S. C., Wettenhall, R. E., & Thomas, G.
- Affiliations
- Friedrich Miescher Institute (CH)
- Venue
- The EMBO Journal
- Published
- 1995-11-01
- Volume
- 14
- Issue
- 21
- Pages
- 5279-5287
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC394637
- OpenAlex
- W1585385265
- Cited by
- 443
- References
- 0
- Citations by year
- 2026: 2 · 2025: 5 · 2024: 3 · 2023: 7 · 2022: 5 · 2021: 10 · 2020: 6 · 2019: 10 · 2018: 5 · 2017: 5 · 2016: 9 · 2015: 12 · 2014: 19 · 2013: 12 · 2012: 17
- FWCI
- 6.70
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The EMBO Journal (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Medical Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › PI3K/AKT/mTOR signaling in cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolism, Diabetes, and Cancer
- Keywords
- p70S6K, rapamycin, mTOR, phosphorylation sites, hydrophobic domains, kinase dephosphorylation, constitutive kinase activity, rapamycin resistance
- MeSH terms
- Amino Acid Sequence, Animals, Antifungal Agents, Enzyme Activation, Humans, Molecular Sequence Data, Phosphorylation, Polyenes, Mutagenesis, Site-Directed, 3T3 Cells, Conserved Sequence, Protein Serine-Threonine Kinases, Sequence Analysis, Ribosomal Protein S6 Kinases, Sirolimus, Mice
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