Abstract
Stimulation of AMP-activated protein kinase (AMPK) in skeletal muscle has been correlated with an increase in glucose transport. Here, we demonstrate that adenoviral-mediated expression of a constitutively active mutant of AMPK alpha leads to activation of glucose transport in a skeletal-muscle cell line, similar to that seen following treatment with 5-amino-imidazolecarboxamide (AICA) riboside, hyperosmotic stress or insulin. In contrast, expression of a dominant-negative form of AMPK blocked the stimulation of glucose transport by both AICA riboside and hyperosmotic stress, but was without effect on either insulin or phorbol-ester-stimulated transport. These results demonstrate that activation of AMPK is sufficient for stimulation of glucose uptake into muscle cells, and is a necessary component of the AICA riboside- and hyperosmotic-stress-induced pathway leading to increased glucose uptake. On the other hand, AMPK is not required in the insulin- or phorbol-ester-mediated pathways. Long-term (5 days) expression of the constitutively active AMPK mutant increased protein expression of GLUT1, GLUT4 and hexokinase II, consistent with previous reports on the chronic treatment of rats with AICA riboside. Expression of constitutively active AMPK had no detectable effect on p38 mitogen-activated protein kinase levels, although interestingly the level of protein kinase B was decreased. These results demonstrate that long-term activation of AMPK is sufficient to cause increased expression of specific proteins in muscle. Our results add further support to the hypothesis that long-term activation of AMPK is involved in the adaptive response of muscle to exercise training.
Full Text
The Full Text of this article is available as a PDF (240.6 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Cohen P. The search for physiological substrates of MAP and SAP kinases in mammalian cells. Trends Cell Biol. 1997 Sep;7(9):353–361. doi: 10.1016/S0962-8924(97)01105-7. [DOI] [PubMed] [Google Scholar]
- Corton J. M., Gillespie J. G., Hardie D. G. Role of the AMP-activated protein kinase in the cellular stress response. Curr Biol. 1994 Apr 1;4(4):315–324. doi: 10.1016/s0960-9822(00)00070-1. [DOI] [PubMed] [Google Scholar]
- Corton J. M., Gillespie J. G., Hawley S. A., Hardie D. G. 5-aminoimidazole-4-carboxamide ribonucleoside. A specific method for activating AMP-activated protein kinase in intact cells? Eur J Biochem. 1995 Apr 15;229(2):558–565. doi: 10.1111/j.1432-1033.1995.tb20498.x. [DOI] [PubMed] [Google Scholar]
- Crute B. E., Seefeld K., Gamble J., Kemp B. E., Witters L. A. Functional domains of the alpha1 catalytic subunit of the AMP-activated protein kinase. J Biol Chem. 1998 Dec 25;273(52):35347–35354. doi: 10.1074/jbc.273.52.35347. [DOI] [PubMed] [Google Scholar]
- Dela F., Ploug T., Handberg A., Petersen L. N., Larsen J. J., Mikines K. J., Galbo H. Physical training increases muscle GLUT4 protein and mRNA in patients with NIDDM. Diabetes. 1994 Jul;43(7):862–865. doi: 10.2337/diab.43.7.862. [DOI] [PubMed] [Google Scholar]
- Foretz M., Carling D., Guichard C., Ferré P., Foufelle F. AMP-activated protein kinase inhibits the glucose-activated expression of fatty acid synthase gene in rat hepatocytes. J Biol Chem. 1998 Jun 12;273(24):14767–14771. doi: 10.1074/jbc.273.24.14767. [DOI] [PubMed] [Google Scholar]
- Forsayeth J., Gould M. K. Effects of hyperosmolarity on basal and insulin-stimulated muscle sugar transport. Am J Physiol. 1981 Mar;240(3):E263–E267. doi: 10.1152/ajpendo.1981.240.3.E263. [DOI] [PubMed] [Google Scholar]
- Fryer L. G., Hajduch E., Rencurel F., Salt I. P., Hundal H. S., Hardie D. G., Carling D. Activation of glucose transport by AMP-activated protein kinase via stimulation of nitric oxide synthase. Diabetes. 2000 Dec;49(12):1978–1985. doi: 10.2337/diabetes.49.12.1978. [DOI] [PubMed] [Google Scholar]
- Hardie D. G., Carling D., Carlson M. The AMP-activated/SNF1 protein kinase subfamily: metabolic sensors of the eukaryotic cell? Annu Rev Biochem. 1998;67:821–855. doi: 10.1146/annurev.biochem.67.1.821. [DOI] [PubMed] [Google Scholar]
- Hardie D. G., Carling D. The AMP-activated protein kinase--fuel gauge of the mammalian cell? Eur J Biochem. 1997 Jun 1;246(2):259–273. doi: 10.1111/j.1432-1033.1997.00259.x. [DOI] [PubMed] [Google Scholar]
- Hardie D. G., Salt I. P., Hawley S. A., Davies S. P. AMP-activated protein kinase: an ultrasensitive system for monitoring cellular energy charge. Biochem J. 1999 Mar 15;338(Pt 3):717–722. [PMC free article] [PubMed] [Google Scholar]
- Hashimoto M., Hatanaka Y., Yang J., Dhesi J., Holman G. D. Synthesis of biotinylated bis(D-glucose) derivatives for glucose transporter photoaffinity labelling. Carbohydr Res. 2001 Mar 22;331(2):119–127. doi: 10.1016/s0008-6215(01)00025-8. [DOI] [PubMed] [Google Scholar]
- Hawley S. A., Davison M., Woods A., Davies S. P., Beri R. K., Carling D., Hardie D. G. Characterization of the AMP-activated protein kinase kinase from rat liver and identification of threonine 172 as the major site at which it phosphorylates AMP-activated protein kinase. J Biol Chem. 1996 Nov 1;271(44):27879–27887. doi: 10.1074/jbc.271.44.27879. [DOI] [PubMed] [Google Scholar]
- Hawley S. A., Selbert M. A., Goldstein E. G., Edelman A. M., Carling D., Hardie D. G. 5'-AMP activates the AMP-activated protein kinase cascade, and Ca2+/calmodulin activates the calmodulin-dependent protein kinase I cascade, via three independent mechanisms. J Biol Chem. 1995 Nov 10;270(45):27186–27191. doi: 10.1074/jbc.270.45.27186. [DOI] [PubMed] [Google Scholar]
- Hayashi T., Hirshman M. F., Fujii N., Habinowski S. A., Witters L. A., Goodyear L. J. Metabolic stress and altered glucose transport: activation of AMP-activated protein kinase as a unifying coupling mechanism. Diabetes. 2000 Apr;49(4):527–531. doi: 10.2337/diabetes.49.4.527. [DOI] [PubMed] [Google Scholar]
- Hayashi T., Hirshman M. F., Kurth E. J., Winder W. W., Goodyear L. J. Evidence for 5' AMP-activated protein kinase mediation of the effect of muscle contraction on glucose transport. Diabetes. 1998 Aug;47(8):1369–1373. doi: 10.2337/diab.47.8.1369. [DOI] [PubMed] [Google Scholar]
- Hayashi T., Wojtaszewski J. F., Goodyear L. J. Exercise regulation of glucose transport in skeletal muscle. Am J Physiol. 1997 Dec;273(6 Pt 1):E1039–E1051. doi: 10.1152/ajpendo.1997.273.6.E1039. [DOI] [PubMed] [Google Scholar]
- Holmes B. F., Kurth-Kraczek E. J., Winder W. W. Chronic activation of 5'-AMP-activated protein kinase increases GLUT-4, hexokinase, and glycogen in muscle. J Appl Physiol (1985) 1999 Nov;87(5):1990–1995. doi: 10.1152/jappl.1999.87.5.1990. [DOI] [PubMed] [Google Scholar]
- Johnson L. N., Noble M. E., Owen D. J. Active and inactive protein kinases: structural basis for regulation. Cell. 1996 Apr 19;85(2):149–158. doi: 10.1016/s0092-8674(00)81092-2. [DOI] [PubMed] [Google Scholar]
- Kahn B. B. Type 2 diabetes: when insulin secretion fails to compensate for insulin resistance. Cell. 1998 Mar 6;92(5):593–596. doi: 10.1016/s0092-8674(00)81125-3. [DOI] [PubMed] [Google Scholar]
- Kemp B. E., Mitchelhill K. I., Stapleton D., Michell B. J., Chen Z. P., Witters L. A. Dealing with energy demand: the AMP-activated protein kinase. Trends Biochem Sci. 1999 Jan;24(1):22–25. doi: 10.1016/s0968-0004(98)01340-1. [DOI] [PubMed] [Google Scholar]
- Khayat Z. A., Tsakiridis T., Ueyama A., Somwar R., Ebina Y., Klip A. Rapid stimulation of glucose transport by mitochondrial uncoupling depends in part on cytosolic Ca2+ and cPKC. Am J Physiol. 1998 Dec;275(6 Pt 1):C1487–C1497. doi: 10.1152/ajpcell.1998.275.6.C1487. [DOI] [PubMed] [Google Scholar]
- Kitzmann M., Carnac G., Vandromme M., Primig M., Lamb N. J., Fernandez A. The muscle regulatory factors MyoD and myf-5 undergo distinct cell cycle-specific expression in muscle cells. J Cell Biol. 1998 Sep 21;142(6):1447–1459. doi: 10.1083/jcb.142.6.1447. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kubo K., Foley J. E. Rate-limiting steps for insulin-mediated glucose uptake into perfused rat hindlimb. Am J Physiol. 1986 Jan;250(1 Pt 1):E100–E102. doi: 10.1152/ajpendo.1986.250.1.E100. [DOI] [PubMed] [Google Scholar]
- Kurth-Kraczek E. J., Hirshman M. F., Goodyear L. J., Winder W. W. 5' AMP-activated protein kinase activation causes GLUT4 translocation in skeletal muscle. Diabetes. 1999 Aug;48(8):1667–1671. doi: 10.2337/diabetes.48.8.1667. [DOI] [PubMed] [Google Scholar]
- Kyriakis J. M., Avruch J. Sounding the alarm: protein kinase cascades activated by stress and inflammation. J Biol Chem. 1996 Oct 4;271(40):24313–24316. doi: 10.1074/jbc.271.40.24313. [DOI] [PubMed] [Google Scholar]
- Leclerc I., Kahn A., Doiron B. The 5'-AMP-activated protein kinase inhibits the transcriptional stimulation by glucose in liver cells, acting through the glucose response complex. FEBS Lett. 1998 Jul 17;431(2):180–184. doi: 10.1016/s0014-5793(98)00745-5. [DOI] [PubMed] [Google Scholar]
- Lochhead P. A., Salt I. P., Walker K. S., Hardie D. G., Sutherland C. 5-aminoimidazole-4-carboxamide riboside mimics the effects of insulin on the expression of the 2 key gluconeogenic genes PEPCK and glucose-6-phosphatase. Diabetes. 2000 Jun;49(6):896–903. doi: 10.2337/diabetes.49.6.896. [DOI] [PubMed] [Google Scholar]
- Lund S., Holman G. D., Schmitz O., Pedersen O. Contraction stimulates translocation of glucose transporter GLUT4 in skeletal muscle through a mechanism distinct from that of insulin. Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):5817–5821. doi: 10.1073/pnas.92.13.5817. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marsin A. S., Bertrand L., Rider M. H., Deprez J., Beauloye C., Vincent M. F., Van den Berghe G., Carling D., Hue L. Phosphorylation and activation of heart PFK-2 by AMPK has a role in the stimulation of glycolysis during ischaemia. Curr Biol. 2000 Oct 19;10(20):1247–1255. doi: 10.1016/s0960-9822(00)00742-9. [DOI] [PubMed] [Google Scholar]
- Merrill G. F., Kurth E. J., Hardie D. G., Winder W. W. AICA riboside increases AMP-activated protein kinase, fatty acid oxidation, and glucose uptake in rat muscle. Am J Physiol. 1997 Dec;273(6 Pt 1):E1107–E1112. doi: 10.1152/ajpendo.1997.273.6.E1107. [DOI] [PubMed] [Google Scholar]
- Mu J., Brozinick J. T., Jr, Valladares O., Bucan M., Birnbaum M. J. A role for AMP-activated protein kinase in contraction- and hypoxia-regulated glucose transport in skeletal muscle. Mol Cell. 2001 May;7(5):1085–1094. doi: 10.1016/s1097-2765(01)00251-9. [DOI] [PubMed] [Google Scholar]
- Neufer P. D., Dohm G. L. Exercise induces a transient increase in transcription of the GLUT-4 gene in skeletal muscle. Am J Physiol. 1993 Dec;265(6 Pt 1):C1597–C1603. doi: 10.1152/ajpcell.1993.265.6.C1597. [DOI] [PubMed] [Google Scholar]
- Ponticos M., Lu Q. L., Morgan J. E., Hardie D. G., Partridge T. A., Carling D. Dual regulation of the AMP-activated protein kinase provides a novel mechanism for the control of creatine kinase in skeletal muscle. EMBO J. 1998 Mar 16;17(6):1688–1699. doi: 10.1093/emboj/17.6.1688. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Qin S., Minami Y., Hibi M., Kurosaki T., Yamamura H. Syk-dependent and -independent signaling cascades in B cells elicited by osmotic and oxidative stress. J Biol Chem. 1997 Jan 24;272(4):2098–2103. doi: 10.1074/jbc.272.4.2098. [DOI] [PubMed] [Google Scholar]
- Ren J. M., Semenkovich C. F., Gulve E. A., Gao J., Holloszy J. O. Exercise induces rapid increases in GLUT4 expression, glucose transport capacity, and insulin-stimulated glycogen storage in muscle. J Biol Chem. 1994 May 20;269(20):14396–14401. [PubMed] [Google Scholar]
- Russell R. R., 3rd, Bergeron R., Shulman G. I., Young L. H. Translocation of myocardial GLUT-4 and increased glucose uptake through activation of AMPK by AICAR. Am J Physiol. 1999 Aug;277(2 Pt 2):H643–H649. doi: 10.1152/ajpheart.1999.277.2.H643. [DOI] [PubMed] [Google Scholar]
- Stein S. C., Woods A., Jones N. A., Davison M. D., Carling D. The regulation of AMP-activated protein kinase by phosphorylation. Biochem J. 2000 Feb 1;345(Pt 3):437–443. [PMC free article] [PubMed] [Google Scholar]
- Sullivan J. E., Brocklehurst K. J., Marley A. E., Carey F., Carling D., Beri R. K. Inhibition of lipolysis and lipogenesis in isolated rat adipocytes with AICAR, a cell-permeable activator of AMP-activated protein kinase. FEBS Lett. 1994 Oct 10;353(1):33–36. doi: 10.1016/0014-5793(94)01006-4. [DOI] [PubMed] [Google Scholar]
- Sullivan J. E., Carey F., Carling D., Beri R. K. Characterisation of 5'-AMP-activated protein kinase in human liver using specific peptide substrates and the effects of 5'-AMP analogues on enzyme activity. Biochem Biophys Res Commun. 1994 May 16;200(3):1551–1556. doi: 10.1006/bbrc.1994.1627. [DOI] [PubMed] [Google Scholar]
- Van der Kaay J., Beck M., Gray A., Downes C. P. Distinct phosphatidylinositol 3-kinase lipid products accumulate upon oxidative and osmotic stress and lead to different cellular responses. J Biol Chem. 1999 Dec 10;274(50):35963–35968. doi: 10.1074/jbc.274.50.35963. [DOI] [PubMed] [Google Scholar]
- Vavvas D., Apazidis A., Saha A. K., Gamble J., Patel A., Kemp B. E., Witters L. A., Ruderman N. B. Contraction-induced changes in acetyl-CoA carboxylase and 5'-AMP-activated kinase in skeletal muscle. J Biol Chem. 1997 May 16;272(20):13255–13261. doi: 10.1074/jbc.272.20.13255. [DOI] [PubMed] [Google Scholar]
- Winder W. W., Hardie D. G. Inactivation of acetyl-CoA carboxylase and activation of AMP-activated protein kinase in muscle during exercise. Am J Physiol. 1996 Feb;270(2 Pt 1):E299–E304. doi: 10.1152/ajpendo.1996.270.2.E299. [DOI] [PubMed] [Google Scholar]
- Winder W. W., Holmes B. F., Rubink D. S., Jensen E. B., Chen M., Holloszy J. O. Activation of AMP-activated protein kinase increases mitochondrial enzymes in skeletal muscle. J Appl Physiol (1985) 2000 Jun;88(6):2219–2226. doi: 10.1152/jappl.2000.88.6.2219. [DOI] [PubMed] [Google Scholar]
- Woods A., Azzout-Marniche D., Foretz M., Stein S. C., Lemarchand P., Ferré P., Foufelle F., Carling D. Characterization of the role of AMP-activated protein kinase in the regulation of glucose-activated gene expression using constitutively active and dominant negative forms of the kinase. Mol Cell Biol. 2000 Sep;20(18):6704–6711. doi: 10.1128/mcb.20.18.6704-6711.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woods A., Cheung P. C., Smith F. C., Davison M. D., Scott J., Beri R. K., Carling D. Characterization of AMP-activated protein kinase beta and gamma subunits. Assembly of the heterotrimeric complex in vitro. J Biol Chem. 1996 Apr 26;271(17):10282–10290. doi: 10.1074/jbc.271.17.10282. [DOI] [PubMed] [Google Scholar]
- Woods A., Salt I., Scott J., Hardie D. G., Carling D. The alpha1 and alpha2 isoforms of the AMP-activated protein kinase have similar activities in rat liver but exhibit differences in substrate specificity in vitro. FEBS Lett. 1996 Nov 18;397(2-3):347–351. doi: 10.1016/s0014-5793(96)01209-4. [DOI] [PubMed] [Google Scholar]
- Yeh J. I., Gulve E. A., Rameh L., Birnbaum M. J. The effects of wortmannin on rat skeletal muscle. Dissociation of signaling pathways for insulin- and contraction-activated hexose transport. J Biol Chem. 1995 Feb 3;270(5):2107–2111. doi: 10.1074/jbc.270.5.2107. [DOI] [PubMed] [Google Scholar]
- Young M. E., Radda G. K., Leighton B. Activation of glycogen phosphorylase and glycogenolysis in rat skeletal muscle by AICAR--an activator of AMP-activated protein kinase. FEBS Lett. 1996 Mar 11;382(1-2):43–47. doi: 10.1016/0014-5793(96)00129-9. [DOI] [PubMed] [Google Scholar]