Control of p70 ribosomal protein S6 kinase and acetyl-CoA carboxylase by AMP-activated protein kinase and protein phosphatases in isolated hepatocytes

Eur J Biochem. 2002 Aug;269(15):3751-9. doi: 10.1046/j.1432-1033.2002.03074.x.

Abstract

Certain amino acids, like glutamine and leucine, induce an anabolic response in liver. They activate p70 ribosomal protein S6 kinase (p70S6K) and acetyl-CoA carboxylase (ACC) involved in protein and fatty acids synthesis, respectively. In contrast, the AMP-activated protein kinase (AMPK), which senses the energy state of the cell and becomes activated under metabolic stress, inactivates by phosphorylation key enzymes in biosynthetic pathways thereby conserving ATP. In this paper, we studied the effect of AMPK activation and of protein phosphatase inhibitors, on the amino-acid-induced activation of p70S6K and ACC in hepatocytes in suspension. AMPK was activated under anoxic conditions or by incubation with 5-aminoimidazole-4-carboxyamide ribonucleoside (AICAr) or oligomycin, an inhibitor of mitochondrial oxidative phosphorylation. Incubation of hepatocytes with amino acids activated p70S6K via multiple phosphorylation. It also activated ACC by a phosphatase-dependent mechanism but did not modify AMPK activation. Conversely, the amino-acid-induced activation of both ACC and p70S6K was blocked or reversed when AMPK was activated. This AMPK activation increased Ser79 phosphorylation in ACC but decreased Thr389 phosphorylation in p70S6K. Protein phosphatase inhibitors prevented p70S6K activation when added prior to the incubation with amino acids, whereas they enhanced p70S6K activation when added after the preincubation with amino acids. It is concluded that (a) AMPK blocks amino-acid-induced activation of ACC and p70S6K, directly by phosphorylating Ser79 in ACC, and indirectly by inhibiting p70S6K phosphorylation, and (b) both activation and inhibition of protein phosphatases are involved in the activation of p70S6K by amino acids. p70S6K adds to an increasing list of targets of AMPK in agreement with the inhibition of energy-consuming biosynthetic pathways.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases
  • Acetyl-CoA Carboxylase / drug effects
  • Acetyl-CoA Carboxylase / metabolism*
  • Aminoimidazole Carboxamide / analogs & derivatives*
  • Aminoimidazole Carboxamide / pharmacology
  • Animals
  • Cells, Cultured
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology
  • Glutamine / metabolism
  • Glutamine / pharmacology
  • Hepatocytes / drug effects
  • Hepatocytes / metabolism*
  • Male
  • Multienzyme Complexes / drug effects
  • Multienzyme Complexes / metabolism*
  • Okadaic Acid / pharmacology
  • Oligomycins / pharmacology
  • Oxazoles / pharmacology
  • Phosphoprotein Phosphatases / antagonists & inhibitors
  • Phosphoprotein Phosphatases / drug effects
  • Phosphoprotein Phosphatases / metabolism*
  • Phosphorylation
  • Protein-Serine-Threonine Kinases / drug effects
  • Protein-Serine-Threonine Kinases / metabolism*
  • Rats
  • Rats, Wistar
  • Ribonucleotides / pharmacology
  • Ribosomal Protein S6 Kinases / drug effects
  • Ribosomal Protein S6 Kinases / metabolism*
  • Serine / metabolism
  • Sirolimus / pharmacology

Substances

  • Enzyme Inhibitors
  • Multienzyme Complexes
  • Oligomycins
  • Oxazoles
  • Ribonucleotides
  • Glutamine
  • Okadaic Acid
  • Aminoimidazole Carboxamide
  • Serine
  • calyculin A
  • Protein-Serine-Threonine Kinases
  • Ribosomal Protein S6 Kinases
  • AMP-Activated Protein Kinases
  • Phosphoprotein Phosphatases
  • Acetyl-CoA Carboxylase
  • AICA ribonucleotide
  • Sirolimus