A role for AMPK in increased insulin action after serum starvation.
Serum starvation is a common cell culture procedure for increasing cellular response to insulin, though the mechanism for the serum starvation effect is not understood. We hypothesized that factors known to potentiate insulin action [e.g., AMP-activated protein kinase (AMPK) and p38] or to be involved in insulin signaling leading to glucose transport [e.g., Akt, PKCζ, AS160, and ataxia telangiectasia mutated (ATM)] would be phosphorylated during serum starvation and would be responsible for increased insulin action after serum starvation. L6 myotubes were incubated in serum-containing or serum-free medium for 3 h. Levels of phosphorylated AMPK, Akt, and ATM were greater in serum-starved cells than in control cells. Serum starvation did not affect p38, PKCζ, or AS160 phosphorylation or insulin-stimulated Akt or AS160 phosphorylation. Insulin had no effect on glucose transport in control cells but caused an increase in glucose uptake for serum-starved cells that was preventable by compound C (an AMPK inhibitor), by expression of dominant negative AMPK (AMPK-DN), and by KU55933 (an ATM inhibitor). ATM protein levels increased during serum starvation, and this increase in ATM was prevented by compound C and AMPK-DN. Thus, it appears that AMPK is required for the serum starvation-related increase in insulin-stimulated glucose transport, with ATM as a possible downstream effector.
Ching
James Kain
JK
Department of Biology, Saint Louis University, St. Louis, Missouri 63103, USA.
Rajguru
Pooja
P
Marupudi
Nandhini
N
Banerjee
Sankha
S
Fisher
Jonathan S
JS
D055372:AMP-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D064007:Ataxia Telangiectasia Mutated Proteins; D001692:Biological Transport / Q000502:physiology; D018929:Cell Culture Techniques; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D007328:Insulin / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D013217:Starvation / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D007328:Insulin; D007527:Isoenzymes; D011720:Pyrazoles; D011743:Pyrimidines; D025521:Tumor Suppressor Proteins; C516138:dorsomorphin; D064007:Ataxia Telangiectasia Mutated Proteins; C576325:Atm protein, mouse; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases; D005947:Glucose
Am J Physiol Cell Physiol. 2007 Jan;292(1):C564-72
16870829
Anal Biochem. 2010 Feb 15;397(2):233-40
19854150
Cell Metab. 2005 Oct;2(4):263-72
16213228
Physiology (Bethesda). 2006 Feb;21:48-60
16443822
Cell Signal. 2008 Aug;20(8):1555-63
18534819
Biochem J. 2009 Apr 15;419(2):475-84
19140804
Diabetes. 2002 Oct;51(10):2886-94
12351423
J Appl Physiol (1985). 2005 Sep;99(3):877-83
15860681
J Biol Chem. 2008 Apr 4;283(14):9187-95
18258599
Cell Metab. 2006 Nov;4(5):377-89
17084711
Endocrinology. 2005 Dec;146(12):5071-8
16150904
J Biol Chem. 2003 Aug 15;278(33):31000-6
12788940
Am J Physiol Endocrinol Metab. 2002 Jan;282(1):E18-23
11739078
J Biol Chem. 2004 Jan 9;279(2):1070-9
14573616
J Biol Chem. 2002 Jul 12;277(28):25226-32
11994296
Am J Physiol. 1984 Sep;247(3 Pt 1):E291-6
6383069
Cellscience. 2006 Jan 28;2(3):68-81
18049717
J Clin Invest. 2001 Oct;108(8):1167-74
11602624
Cancer Res. 2004 Dec 15;64(24):9152-9
15604286
Eur J Biochem. 1995 Apr 15;229(2):558-65
7744080
Diabetes. 2006 Jul;55(7):2051-8
16804075
Mol Cell Biochem. 2007 Dec;306(1-2):239-45
17786544
Am J Physiol Endocrinol Metab. 2007 Jul;293(1):E70-4
17356010
Physiology (Bethesda). 2005 Aug;20:271-84
16024515
Diabetes. 2000 Mar;49(3):325-31
10868952
Biochem Biophys Res Commun. 2004 Nov 19;324(3):986-92
15485651
Diabetes. 2007 Feb;56(2):414-23
17259386
J Biol Chem. 2002 Sep 6;277(36):32552-7
12107173
Cell. 1996 Jul 12;86(1):159-71
8689683
J Biol Chem. 2006 Oct 20;281(42):31478-85
16935857
Am J Physiol Endocrinol Metab. 2008 Jul;295(1):E29-37
18477703
Appl Physiol Nutr Metab. 2007 Jun;32(3):557-66
17510697
J Biol Chem. 2005 Feb 11;280(6):4029-36
15546863
Acta Physiol (Oxf). 2010 Apr;198(4):465-75
20003097
Diabetes. 2006 Jul;55(7):2067-76
16804077
Proc Natl Acad Sci U S A. 2008 Jun 3;105(22):7785-90
18505846
Mol Cell. 2001 May;7(5):1085-94
11389854
J Clin Invest. 2003 Jan;111(1):91-8
12511592
J Biol Chem. 2008 Oct 3;283(40):27208-19
18650435
Mol Biol Cell. 2006 Oct;17(10):4484-93
16914513
Am J Physiol Endocrinol Metab. 2005 Apr;288(4):E782-8
15585585