A decrease in cellular energy status stimulates PERK-dependent eIF2alpha phosphorylation and regulates protein synthesis in pancreatic beta-cells.
In the present study, we demonstrate that, in pancreatic beta-cells, eIF2alpha (eukaryotic initiation factor 2alpha) phosphorylation in response to a decrease in glucose concentration is primarily mediated by the activation of PERK [PKR (protein kinase RNA activated)-like endoplasmic reticulum kinase]. We provide evidence that this increase in PERK activity is evoked by a decrease in the energy status of the cell via a potentially novel mechanism that is independent of IRE1 (inositol requiring enzyme 1) activation and the accumulation of unfolded nascent proteins within the endoplasmic reticulum. The inhibition of eIF2alpha phosphorylation in glucose-deprived cells by the overexpression of dominant-negative PERK or an N-terminal truncation mutant of GADD34 (growth-arrest and DNA-damage-inducible protein 34) leads to a 53% increase in the rate of total protein synthesis. Polysome analysis revealed that this coincides with an increase in the amplitude but not the number of ribosomes per mRNA, indicating that eIF2alpha dephosphorylation mobilizes hitherto untranslated mRNAs on to polysomes. In summary, we show that PERK is activated at low glucose concentrations in response to a decrease in energy status and that this plays an important role in glucose-regulated protein synthesis in pancreatic beta-cells.
Department of Cell Physiology and Pharmacology, Faculty of Medicine and Biological Sciences, The Henry Wellcome Building, University of Leicester, University Road, Leicester LE1 9HN, UK.
Edith
E
Gomez
Mike L
ML
Powell
Alan
A
Bevington
Terence P
TP
Herbert
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004734:Energy Metabolism; D004789:Enzyme Activation; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism*; D020868:Gene Silencing; D005947:Glucose / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D034741:RNA, Small Interfering; D019892:eIF-2 Kinase / Q000235:genetics / Q000378:metabolism*
D000255:Adenosine Triphosphate; D017931:DNA Primers; D015852:Eukaryotic Initiation Factor-2; D005947:Glucose; D034741:RNA, Small Interfering; D019892:eIF-2 Kinase