Role for AMP-activated protein kinase in glucose-stimulated insulin secretion and preproinsulin gene expression.
AMP-activated protein kinase (AMPK) has recently been implicated in the control of preproinsulin gene expression in pancreatic islet beta-cells [da Silva Xavier, Leclerc, Salt, Doiron, Hardie, Kahn and Rutter (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 4023-4028]. Using pharmacological and molecular strategies to regulate AMPK activity in rat islets and clonal MIN6 beta-cells, we show here that the effects of AMPK are exerted largely upstream of insulin release. Thus forced increases in AMPK activity achieved pharmacologically with 5-amino-4-imidazolecarboxamide riboside (AICAR), or by adenoviral overexpression of a truncated, constitutively active form of the enzyme (AMPK alpha 1.T(172)D), blocked glucose-stimulated insulin secretion. In MIN6 cells, activation of AMPK suppressed glucose metabolism, as assessed by changes in total, cytosolic or mitochondrial [ATP] and NAD(P)H, and reduced increases in intracellular [Ca(2+)] caused by either glucose or tolbutamide. By contrast, inactivation of AMPK by expression of a dominant-negative form of the enzyme mutated in the catalytic site (AMPK alpha 1.D(157)A) did not affect glucose-stimulated increases in [ATP], NAD(P)H or intracellular [Ca(2+)], but led to the unregulated release of insulin. These results indicate that inhibition of AMPK by glucose is essential for the activation of insulin secretion by the sugar, and may contribute to the transcriptional stimulation of the preproinsulin gene. Modulation of AMPK activity in the beta-cell may thus represent a novel therapeutic strategy for the treatment of type 2 diabetes mellitus.
da Silva Xavier
Gabriela
G
Henry Wellcome Laboratories of Integrated Cell Signalling and Department of Biochemistry, University Walk, University of Bristol, Bristol BS8 1TD, UK.
Leclerc
Isabelle
I
Varadi
Aniko
A
Tsuboi
Takashi
T
Moule
S Kelly
SK
Rutter
Guy A
GA
D017209:Apoptosis; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D005786:Gene Expression Regulation; D005947:Glucose / Q000494:pharmacology*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D010766:Phosphorylation; D011384:Proinsulin / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics*
D007328:Insulin; D011498:Protein Precursors; C013737:preproinsulin; D011384:Proinsulin; D017868:Cyclic AMP-Dependent Protein Kinases; D005947:Glucose; D002118:Calcium