• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. May 1, 1996; 15(9): 2291–2297.
PMCID: PMC450154

Regulation of translation elongation factor-2 by insulin via a rapamycin-sensitive signalling pathway.

Abstract

It is well established that insulin and serum stimulate gene expression at the level of mRNA translation in animal cells, and previous studies have mainly focused on the initiation process. Here we show that, in Chinese hamster ovary cells expressing the human insulin receptor, insulin causes decreased phosphorylation of elongation factor eEF-2 and that this is associated with stimulation of the rate of peptide-chain elongation. eEF-2 is phosphorylated by a very specific Ca 2+/calmodulin-dependent protein kinase (eEF-2 kinase) causing its complete inactivation. The decrease in eEF-2 phosphorylation induced by insulin reflects a fall in eEF-2 kinase activity. Rapamycin, a macrolide immunosuppressant which blocks the signalling pathway leading to the stimulation of the 70/85 kDa ribosomal protein S6 kinases, substantially blocks the activation of elongation, the fall in eEF-2 phosphorylation and the decrease in eEF-2 kinase activity, suggesting that p7O S6 kinase (p70s6k) and eEF-2 kinase may tie on a common signalling pathway. Wortmannin, an inhibitor of phosphatidylinositide-3-OH kinase, had similar effects. eEF-2 kinase was phosphorylated in vitro by purified p70s6k but this had no significant effect on the in vitro activity of eEF-2 kinase.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.3M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Altmann M, Trachsel H. Regulation of translation initiation and modulation of cellular physiology. Trends Biochem Sci. 1993 Nov;18(11):429–432. [PubMed]
  • Avruch J, Zhang XF, Kyriakis JM. Raf meets Ras: completing the framework of a signal transduction pathway. Trends Biochem Sci. 1994 Jul;19(7):279–283. [PubMed]
  • Brandis JW, Raff RA. Elevation of protein synthesis is a complex response to fertilisation. Nature. 1979 Mar 29;278(5703):467–469. [PubMed]
  • Chung J, Kuo CJ, Crabtree GR, Blenis J. Rapamycin-FKBP specifically blocks growth-dependent activation of and signaling by the 70 kd S6 protein kinases. Cell. 1992 Jun 26;69(7):1227–1236. [PubMed]
  • Cross DA, Alessi DR, Vandenheede JR, McDowell HE, Hundal HS, Cohen P. The inhibition of glycogen synthase kinase-3 by insulin or insulin-like growth factor 1 in the rat skeletal muscle cell line L6 is blocked by wortmannin, but not by rapamycin: evidence that wortmannin blocks activation of the mitogen-activated protein kinase pathway in L6 cells between Ras and Raf. Biochem J. 1994 Oct 1;303(Pt 1):21–26. [PMC free article] [PubMed]
  • Demolle D, Lecomte M, Boutherin-Falson O, Cragoe EJ, Jr, Nairn AC, Boeynaems JM. Amiloride analogs induce the phosphorylation of elongation factor-2 in vascular endothelial cells. Mol Pharmacol. 1990 Jun;37(6):827–832. [PubMed]
  • Dickens M, Chin JE, Roth RA, Ellis L, Denton RM, Tavaré JM. Characterization of insulin-stimulated protein serine/threonine kinases in CHO cells expressing human insulin receptors with point and deletion mutations. Biochem J. 1992 Oct 1;287(Pt 1):201–209. [PMC free article] [PubMed]
  • Ferrari S, Pearson RB, Siegmann M, Kozma SC, Thomas G. The immunosuppressant rapamycin induces inactivation of p70s6k through dephosphorylation of a novel set of sites. J Biol Chem. 1993 Aug 5;268(22):16091–16094. [PubMed]
  • Fingar DC, Hausdorff SF, Blenis J, Birnbaum MJ. Dissociation of pp70 ribosomal protein S6 kinase from insulin-stimulated glucose transport in 3T3-L1 adipocytes. J Biol Chem. 1993 Feb 5;268(4):3005–3008. [PubMed]
  • Fischer I, Arfin SM, Moldave K. Regulation of translation. Analysis of intermediary reactions in protein synthesis in exponentially growing and stationary phase Chinese hamster ovary cells in culture. Biochemistry. 1980 Apr 1;19(7):1417–1425. [PubMed]
  • Grove JR, Banerjee P, Balasubramanyam A, Coffer PJ, Price DJ, Avruch J, Woodgett JR. Cloning and expression of two human p70 S6 kinase polypeptides differing only at their amino termini. Mol Cell Biol. 1991 Nov;11(11):5541–5550. [PMC free article] [PubMed]
  • Gschwendt M, Kittstein W, Mieskes G, Marks F. A type 2A protein phosphatase dephosphorylates the elongation factor 2 and is stimulated by the phorbol ester TPA in mouse epidermis in vivo. FEBS Lett. 1989 Nov 6;257(2):357–360. [PubMed]
  • Han JW, Pearson RB, Dennis PB, Thomas G. Rapamycin, wortmannin, and the methylxanthine SQ20006 inactivate p70s6k by inducing dephosphorylation of the same subset of sites. J Biol Chem. 1995 Sep 8;270(36):21396–21403. [PubMed]
  • Hassell JA, Engelhardt DL. The regulation of protein synthesis in animal cells by serum factors. Biochemistry. 1976 Apr 6;15(7):1375–1381. [PubMed]
  • Hershey JW. Translational control in mammalian cells. Annu Rev Biochem. 1991;60:717–755. [PubMed]
  • Hille MB, Albers AA. Efficiency of protein synthesis after fertilisation of sea urchin eggs. Nature. 1979 Mar 29;278(5703):469–471. [PubMed]
  • Hincke MT, Nairn AC. Phosphorylation of elongation factor 2 during Ca(2+)-mediated secretion from rat parotid acini. Biochem J. 1992 Mar 15;282(Pt 3):877–882. [PMC free article] [PubMed]
  • Kimball SR, Vary TC, Jefferson LS. Regulation of protein synthesis by insulin. Annu Rev Physiol. 1994;56:321–348. [PubMed]
  • Koromilas AE, Lazaris-Karatzas A, Sonenberg N. mRNAs containing extensive secondary structure in their 5' non-coding region translate efficiently in cells overexpressing initiation factor eIF-4E. EMBO J. 1992 Nov;11(11):4153–4158. [PMC free article] [PubMed]
  • Kozma SC, Thomas G. p70s6k/p85s6k: mechanism of activation and role in mitogenesis. Semin Cancer Biol. 1994 Aug;5(4):255–260. [PubMed]
  • Kozma SC, Ferrari S, Thomas G. Unmasking a growth factor/oncogene-activated S6 phosphorylation cascade. Cell Signal. 1989;1(3):219–225. [PubMed]
  • Lane HA, Fernandez A, Lamb NJ, Thomas G. p70s6k function is essential for G1 progression. Nature. 1993 May 13;363(6425):170–172. [PubMed]
  • Lin TA, Kong X, Saltiel AR, Blackshear PJ, Lawrence JC., Jr Control of PHAS-I by insulin in 3T3-L1 adipocytes. Synthesis, degradation, and phosphorylation by a rapamycin-sensitive and mitogen-activated protein kinase-independent pathway. J Biol Chem. 1995 Aug 4;270(31):18531–18538. [PubMed]
  • Mitsui K, Brady M, Palfrey HC, Nairn AC. Purification and characterization of calmodulin-dependent protein kinase III from rabbit reticulocytes and rat pancreas. J Biol Chem. 1993 Jun 25;268(18):13422–13433. [PubMed]
  • Nairn AC, Palfrey HC. Identification of the major Mr 100,000 substrate for calmodulin-dependent protein kinase III in mammalian cells as elongation factor-2. J Biol Chem. 1987 Dec 25;262(36):17299–17303. [PubMed]
  • Nielsen PJ, McConkey EH. Evidence for control of protein synthesis in HeLa cells via the elongation rate. J Cell Physiol. 1980 Sep;104(3):269–281. [PubMed]
  • Palfrey HC. Presence in many mammalian tissues of an identical major cytosolic substrate (Mr 100 000) for calmodulin-dependent protein kinase. FEBS Lett. 1983 Jun 27;157(1):183–190. [PubMed]
  • Palfrey HC, Nairn AC, Muldoon LL, Villereal ML. Rapid activation of calmodulin-dependent protein kinase III in mitogen-stimulated human fibroblasts. Correlation with intracellular Ca2+ transients. J Biol Chem. 1987 Jul 15;262(20):9785–9792. [PubMed]
  • Pause A, Belsham GJ, Gingras AC, Donzé O, Lin TA, Lawrence JC, Jr, Sonenberg N. Insulin-dependent stimulation of protein synthesis by phosphorylation of a regulator of 5'-cap function. Nature. 1994 Oct 27;371(6500):762–767. [PubMed]
  • Price DJ, Grove JR, Calvo V, Avruch J, Bierer BE. Rapamycin-induced inhibition of the 70-kilodalton S6 protein kinase. Science. 1992 Aug 14;257(5072):973–977. [PubMed]
  • Proud CG. Peptide-chain elongation in eukaryotes. Mol Biol Rep. 1994 May;19(3):161–170. [PubMed]
  • Redpath NT. High-resolution one-dimensional polyacrylamide gel isoelectric focusing of various forms of elongation factor-2. Anal Biochem. 1992 May 1;202(2):340–343. [PubMed]
  • Redpath NT, Proud CG. The tumour promoter okadaic acid inhibits reticulocyte-lysate protein synthesis by increasing the net phosphorylation of elongation factor 2. Biochem J. 1989 Aug 15;262(1):69–75. [PMC free article] [PubMed]
  • Redpath NT, Proud CG. Activity of protein phosphatases against initiation factor-2 and elongation factor-2. Biochem J. 1990 Nov 15;272(1):175–180. [PMC free article] [PubMed]
  • Redpath NT, Proud CG. Purification and phosphorylation of elongation factor-2 kinase from rabbit reticulocytes. Eur J Biochem. 1993 Mar 1;212(2):511–520. [PubMed]
  • Redpath NT, Proud CG. Molecular mechanisms in the control of translation by hormones and growth factors. Biochim Biophys Acta. 1994 Jan 13;1220(2):147–162. [PubMed]
  • Ryazanov AG, Natapov PG, Shestakova EA, Severin FF, Spirin AS. Phosphorylation of the elongation factor 2: the fifth Ca2+/calmodulin-dependent system of protein phosphorylation. Biochimie. 1988 May;70(5):619–626. [PubMed]
  • Ryazanov AG, Shestakova EA, Natapov PG. Phosphorylation of elongation factor 2 by EF-2 kinase affects rate of translation. Nature. 1988 Jul 14;334(6178):170–173. [PubMed]
  • Ryazanov AG, Rudkin BB, Spirin AS. Regulation of protein synthesis at the elongation stage. New insights into the control of gene expression in eukaryotes. FEBS Lett. 1991 Jul 22;285(2):170–175. [PubMed]
  • Saito Y, Vandenheede JR, Cohen P. The mechanism by which epidermal growth factor inhibits glycogen synthase kinase 3 in A431 cells. Biochem J. 1994 Oct 1;303(Pt 1):27–31. [PMC free article] [PubMed]
  • Standart N, Jackson R. Translational regulation. Y the message is masked? Curr Biol. 1994 Oct 1;4(10):939–941. [PubMed]
  • Terada N, Lucas JJ, Szepesi A, Franklin RA, Takase K, Gelfand EW. Rapamycin inhibits the phosphorylation of p70 S6 kinase in IL-2 and mitogen-activated human T cells. Biochem Biophys Res Commun. 1992 Aug 14;186(3):1315–1321. [PubMed]
  • Welsh GI, Proud CG. Regulation of protein synthesis in Swiss 3T3 fibroblasts. Rapid activation of the guanine-nucleotide-exchange factor by insulin and growth factors. Biochem J. 1992 May 15;284(Pt 1):19–23. [PMC free article] [PubMed]
  • Welsh GI, Proud CG. Glycogen synthase kinase-3 is rapidly inactivated in response to insulin and phosphorylates eukaryotic initiation factor eIF-2B. Biochem J. 1993 Sep 15;294(Pt 3):625–629. [PMC free article] [PubMed]
  • Welsh GI, Foulstone EJ, Young SW, Tavaré JM, Proud CG. Wortmannin inhibits the effects of insulin and serum on the activities of glycogen synthase kinase-3 and mitogen-activated protein kinase. Biochem J. 1994 Oct 1;303(Pt 1):15–20. [PMC free article] [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...