PERK (EIF2AK3) regulates proinsulin trafficking and quality control in the secretory pathway.
OBJECTIVE
Loss-of-function mutations in Perk (EIF2AK3) result in permanent neonatal diabetes in humans (Wolcott-Rallison Syndrome) and mice. Previously, we found that diabetes associated with Perk deficiency resulted from insufficient proliferation of beta-cells and from defects in insulin secretion. A substantial fraction of PERK-deficient beta-cells display a highly abnormal cellular phenotype characterized by grossly distended endoplasmic reticulum (ER) and retention of proinsulin. We investigated over synthesis, lack of ER-associated degradation (ERAD), and defects in ER to Golgi trafficking as possible causes.


METHODS
ER functions of PERK were investigated in cell culture and mice in which Perk was impaired or gene dosage modulated. The Ins2(+/Akita) mutant mice were used as a model system to test the role of PERK in ERAD.


RESULTS
We report that loss of Perk function does not lead to uncontrolled protein synthesis but impaired ER-to-Golgi anterograde trafficking, retrotranslocation from the ER to the cytoplasm, and proteasomal degradation. PERK was also shown to be required to maintain the integrity of the ER and Golgi and processing of ATF6. Moreover, decreasing Perk dosage surprisingly ameliorates the progression of the Akita mutants toward diabetes.


CONCLUSIONS
PERK is a positive regulator of ERAD and proteasomal activity. Reducing PERK activity ameliorates the progression of diabetes in the Akita mouse, whereas increasing PERK dosage hastens its progression. We speculate that PERK acts as a metabolic sensor in the insulin-secreting beta-cells to modulate the trafficking and quality control of proinsulin in the ER relative to the physiological demands for circulating insulin.
Department of Biology, The Huck Institutes of the Life Sciences, Penn State Institute for Diabetes and Obesity, Pennsylvania State University, University Park, Pennsylvania, USA.
Sounak
S
Gupta
Barbara
B
McGrath
Douglas R
DR
Cavener
D000818:Animals; D003001:Cloning, Molecular; D003920:Diabetes Mellitus / Q000235:genetics; D004721:Endoplasmic Reticulum / Q000502:physiology; D018628:Gene Dosage; D005947:Glucose / Q000494:pharmacology; D006056:Golgi Apparatus / Q000502:physiology; D006801:Humans; D007231:Infant, Newborn; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011384:Proinsulin / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014162:Transfection; D014178:Translocation, Genetic; D019892:eIF-2 Kinase / Q000172:deficiency / Q000235:genetics* / Q000502:physiology*
D005947:Glucose; D007328:Insulin; D011384:Proinsulin; D019892:eIF-2 Kinase; C460640:EIF2AK3 protein, human