Muscle-specific overexpression of wild type and R225Q mutant AMP-activated protein kinase gamma3-subunit differentially regulates glycogen accumulation.
AMP-activated protein kinase (AMPK) is a heterotrimeric complex that works as an energy sensor to integrate nutritional and hormonal signals. The naturally occurring R225Q mutation in the gamma3-subunit in pigs is associated with abnormally high glycogen content in skeletal muscle. Because skeletal muscle accounts for most of the body's glucose uptake, and gamma3 is specifically expressed in skeletal muscle, it is important to understand the underlying mechanism of this mutation in regulating glucose and glycogen metabolism. Using skeletal muscle-specific transgenic mice overexpressing wild type gamma3 (WTgamma3) and R225Q mutant gamma3 (MUTgamma3), we show that both WTgamma3 and MUTgamma3 mice have 1.5- to 2-fold increases in muscle glycogen content. In WTgamma3 mice, increased glycogen content was associated with elevated total glycogen synthase activity and reduced glycogen phosphorylase activity, whereas alterations in activities of these enzymes could not explain elevated glycogen in MUTgamma3 mice. Basal, 5-aminoimidazole-AICAR- and phenformin-stimulated AMPKalpha2 isoform-specific activities were decreased only in MUTgamma3 mice. Basal rates of 2-DG glucose uptake were decreased in both WTgamma3 and MUTgamma3 mice. However, AICAR- and phenformin-stimulated 2-DG glucose uptake were blunted only in MUTgamma3 mice. In conclusion, expression of either wild type or mutant gamma3-subunit of AMPK results in increased glycogen concentrations in muscle, but the mechanisms underlying this alteration appear to be different. Furthermore, mutation of the gamma3-subunit is associated with decreases in AMPKalpha2 isoform-specific activity and impairment in AICAR- and phenformin-stimulated skeletal muscle glucose uptake.
Yu
Haiyan
H
Joslin Diabetes Center, Boston, MA 02215, USA.
Hirshman
Michael F
MF
Fujii
Nobuharu
N
Pomerleau
Jason M
JM
Peter
Lauren E
LE
Goodyear
Laurie J
LJ
D055372:AMP-Activated Protein Kinases; D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D003847:Deoxyglucose / Q000378:metabolism; D005260:Female; D015870:Gene Expression / Q000235:genetics; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006003:Glycogen / Q000378:metabolism*; D024981:Glycogen Phosphorylase / Q000378:metabolism; D006006:Glycogen Synthase / Q000378:metabolism; D007004:Hypoglycemic Agents / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009097:Multienzyme Complexes; D018482:Muscle, Skeletal / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D009206:Myocardium / Q000378:metabolism; D010629:Phenformin / Q000494:pharmacology; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases; D012265:Ribonucleotides / Q000494:pharmacology
D001786:Blood Glucose; D007004:Hypoglycemic Agents; D007328:Insulin; D007527:Isoenzymes; D009097:Multienzyme Complexes; D012265:Ribonucleotides; D000620:Aminoimidazole Carboxamide; D006003:Glycogen; D003847:Deoxyglucose; D010629:Phenformin; D024981:Glycogen Phosphorylase; D006006:Glycogen Synthase; D011494:Protein Kinases; C526221:Prkag3 protein, mouse; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases; C031143:AICA ribonucleotide; D005947:Glucose