Multiple independent inputs are required for activation of the p70 S6 kinase - PubMed (original) (raw)
Multiple independent inputs are required for activation of the p70 S6 kinase
Q P Weng et al. Mol Cell Biol. 1995 May.
Abstract
Previous studies have shown that the noncatalytic carboxy-terminal tail of the p70 S6 kinase (amino acids 422 to 525) contains an autoinhibitory pseudosubstrate domain that is phosphorylated in situ during activation and in vitro by mitogen-activated protein kinases. The present study shows that a recombinant p70 deleted of the carboxy-terminal tail (p70 delta CT104) nevertheless exhibits a basal and serum-stimulated 40S kinase activity and susceptibility to inhibition by wortmannin very similar to those of the parent, full-length p70 kinase. Carboxy-terminal deletion reduces the extent of maximal inhibition produced by rapamycin, from > 95% in the full-length p70 to 60 to 80% in p70 delta CT104, without altering the sensitivity to rapamycin inhibition (50% inhibitory concentration of 2 nM). Serum activation of p70 delta CT104, as with the parent, full-length p70, is accompanied by an increase in 32P content (about twofold) in situ and a slowing in electrophoretic mobility; both modifications are inhibited by pretreatment with wortmannin or rapamycin. 32P-peptide maps of p70 delta CT104 show multisite phosphorylation, and wortmannin and rapamycin appear to cause preferential dephosphorylation of the same subset of sites. Thus, it is likely that activation of the kinase requires phosphorylation of p70 at sites in addition to those previously identified in the carboxy-terminal tail. Evidence that the carboxy-terminal tail actually functions as a potent intramolecular inhibitor of kinase activity in situ is uncovered by deletion of a short acidic segment (amino acids 29 to 46) from the p70 amino-terminal noncatalytic region. Deletion of amino acids 29 to 46 causes a >95% inhibition of p70 activity despite continue phosphorylation of the carboxy-terminal tail in situ; additional deletion of the carboxy-terminal tail (yielding p70 delta 29-46/ delta CT104) increases activity 10-fold, to a level approaching that of p70 delta CT104. Deletion of residues 29 to 46 also abolishes completely the sensitivity of p70 to inhibition by rapamycin but does not alter the susceptibility to activation by serum of inhibition by wortmannin. Although the mechanisms underlying the effects of the delta 29-46 deletion are not known, they are not attributable to loss of the major in situ p70 phosphorylation site at Ser-40. Thus, activation of the p70 S6 kinase involves multiple, independent inputs directed at different domains of the p70 polypeptide. Disinhibition from the carboxy-terminal tail requires, in addition to its multisite phosphorylation, an activating input dependent on the presence of amino acids 29 to 46; this p70-activating input may be the same as that inhibited by rapamycin but is distinct from that arising from the wortmannin-inhibitable phosphatidylinositol 3-kinase. In addition, as exemplified by the rapamycin-resistant but mitogen- and wortmannin-sensitive p70 delta 29-46/ delta CT104 mutant, a further activating input, which probably involves site-specific phosphorylation in the segment between amino acids 46 to 421, is necessary.
Similar articles
- Phosphatidylinositol 3-kinase signals activation of p70 S6 kinase in situ through site-specific p70 phosphorylation.
Weng QP, Andrabi K, Klippel A, Kozlowski MT, Williams LT, Avruch J. Weng QP, et al. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5744-8. doi: 10.1073/pnas.92.12.5744. Proc Natl Acad Sci U S A. 1995. PMID: 7777579 Free PMC article. - The principal rapamycin-sensitive p70(s6k) phosphorylation sites, T-229 and T-389, are differentially regulated by rapamycin-insensitive kinase kinases.
Dennis PB, Pullen N, Kozma SC, Thomas G. Dennis PB, et al. Mol Cell Biol. 1996 Nov;16(11):6242-51. doi: 10.1128/MCB.16.11.6242. Mol Cell Biol. 1996. PMID: 8887654 Free PMC article. - Regulation of the p70 S6 kinase by phosphorylation in vivo. Analysis using site-specific anti-phosphopeptide antibodies.
Weng QP, Kozlowski M, Belham C, Zhang A, Comb MJ, Avruch J. Weng QP, et al. J Biol Chem. 1998 Jun 26;273(26):16621-9. doi: 10.1074/jbc.273.26.16621. J Biol Chem. 1998. PMID: 9632736 - Multiple signaling pathways involved in the metabolic effects of insulin.
Moule SK, Denton RM. Moule SK, et al. Am J Cardiol. 1997 Aug 4;80(3A):41A-49A. doi: 10.1016/s0002-9149(97)00457-8. Am J Cardiol. 1997. PMID: 9293955 Review. - TOR action in mammalian cells and in Caenorhabditis elegans.
Long X, Müller F, Avruch J. Long X, et al. Curr Top Microbiol Immunol. 2004;279:115-38. doi: 10.1007/978-3-642-18930-2_8. Curr Top Microbiol Immunol. 2004. PMID: 14560955 Review.
Cited by
- Single-cell systems pharmacology identifies development-driven drug response and combination therapy in B cell acute lymphoblastic leukemia.
Huang X, Li Y, Zhang J, Yan L, Zhao H, Ding L, Bhatara S, Yang X, Yoshimura S, Yang W, Karol SE, Inaba H, Mullighan C, Litzow M, Zhu X, Zhang Y, Stock W, Jain N, Jabbour E, Kornblau SM, Konopleva M, Pui CH, Paietta E, Evans W, Yu J, Yang JJ. Huang X, et al. Cancer Cell. 2024 Apr 8;42(4):552-567.e6. doi: 10.1016/j.ccell.2024.03.003. Cancer Cell. 2024. PMID: 38593781 Free PMC article. - Understanding the Polyamine and mTOR Pathway Interaction in Breast Cancer Cell Growth.
Akinyele O, Wallace HM. Akinyele O, et al. Med Sci (Basel). 2022 Sep 10;10(3):51. doi: 10.3390/medsci10030051. Med Sci (Basel). 2022. PMID: 36135836 Free PMC article. - Deacetylation of S6 kinase promotes high glucose-induced glomerular mesangial cell hypertrophy and matrix protein accumulation.
Das F, Maity S, Ghosh-Choudhury N, Kasinath BS, Ghosh Choudhury G. Das F, et al. J Biol Chem. 2019 Jun 14;294(24):9440-9460. doi: 10.1074/jbc.RA118.007023. Epub 2019 Apr 26. J Biol Chem. 2019. PMID: 31028173 Free PMC article. - Increased mTORC1 activation in salivary gland B cells and T cells from patients with Sjögren's syndrome: mTOR inhibition as a novel therapeutic strategy to halt immunopathology?
Blokland SLM, Hillen MR, Wichers CGK, Zimmermann M, Kruize AA, Radstake TRDJ, Broen JCA, van Roon JAG. Blokland SLM, et al. RMD Open. 2019 Jan 11;5(1):e000701. doi: 10.1136/rmdopen-2018-000701. eCollection 2019. RMD Open. 2019. PMID: 30713717 Free PMC article. No abstract available. - CD44-mediated activation of α5β1-integrin, cortactin and paxillin signaling underpins adhesion of basal-like breast cancer cells to endothelium and fibronectin-enriched matrices.
McFarlane S, McFarlane C, Montgomery N, Hill A, Waugh DJ. McFarlane S, et al. Oncotarget. 2015 Nov 3;6(34):36762-73. doi: 10.18632/oncotarget.5461. Oncotarget. 2015. PMID: 26447611 Free PMC article.
References
- J Biol Chem. 1989 Aug 15;264(23):13825-33 - PubMed
- Nature. 1988 Aug 25;334(6184):715-8 - PubMed
- J Biol Chem. 1991 Apr 5;266(10):6007-10 - PubMed
- J Biol Chem. 1991 Sep 5;266(25):16281-4 - PubMed
- Mol Cell Biol. 1991 Nov;11(11):5541-50 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials