TY - JOUR
T1 - The inducible beta 5i proteasome subunit contributes to proinsulin degradation in GRP94-deficient beta-cells and is overexpressed in type 2 diabetes pancreatic islets
AU - Khilji, Muhammad Saad
AU - Bresson, Sophie Emilie
AU - Verstappen, Danielle
AU - Pihl, Celina
AU - Andersen, Phillip Alexander Keller
AU - Agergaard, Jette Bach
AU - Dahlby, Tina
AU - Bryde, Tenna Holgersen
AU - Klindt, Kristian
AU - Nielsen, Christian Kronborg
AU - Walentinsson, Anna
AU - Zivkovic, Dusan
AU - Bousquet, Marie-Pierre
AU - Tyrberg, Bjorn
AU - Richardson, Sarah J.
AU - Morgan, Noel G.
AU - Mandrup-Poulsen, Thomas
AU - Marzec, Michal Tomasz
PY - 2020
Y1 - 2020
N2 - Proinsulin is a misfolding-prone protein, and its efficient breakdown is critical when beta-cells are confronted with high-insulin biosynthetic demands, to prevent endoplasmic reticulum stress, a key trigger of secretory dysfunction and, if uncompensated. apoptosis. Proinsulin degradation is thought to be performed by the constitutively expressed standard proteasome, while the roles of other proteasomes are unknown. We recently demonstrated that deficiency of the proinsulin chaperone glucose-regulated protein 94 (GRP94) causes impaired proinsulin handling and defective insulin secretion associated with a compensated endoplasmic reticulum stress response. Taking advantage of this model of restricted folding capacity, we investigated the role of different proteasomes in proinsulin degradation. reasoning that insulin secretory dynamics require an inducible protein degradation system. We show that the expression of only one enzymatically active proteasome subunit, namely, the inducible beta 5i-subunit, was increased in GRP94 CRISPR/Cas9 knockout (KO) cells. Additionally, the level of beta 5i-containing intermediate proteasomes was significantly increased in these cells, as was beta 5i-related chymotrypsin-like activity. Moreover, proinsulin levels were restored in GRP94 KO upon beta 5i small interfering RNA-mediated knockdown. Finally, the fraction of beta-cells expressing the beta 5i-subunit is increased in human islets from type 2 diabetes patients. We conclude that beta 5i is an inducible proteasome subunit dedicated to the degradation of mishandled proinsulin.
AB - Proinsulin is a misfolding-prone protein, and its efficient breakdown is critical when beta-cells are confronted with high-insulin biosynthetic demands, to prevent endoplasmic reticulum stress, a key trigger of secretory dysfunction and, if uncompensated. apoptosis. Proinsulin degradation is thought to be performed by the constitutively expressed standard proteasome, while the roles of other proteasomes are unknown. We recently demonstrated that deficiency of the proinsulin chaperone glucose-regulated protein 94 (GRP94) causes impaired proinsulin handling and defective insulin secretion associated with a compensated endoplasmic reticulum stress response. Taking advantage of this model of restricted folding capacity, we investigated the role of different proteasomes in proinsulin degradation. reasoning that insulin secretory dynamics require an inducible protein degradation system. We show that the expression of only one enzymatically active proteasome subunit, namely, the inducible beta 5i-subunit, was increased in GRP94 CRISPR/Cas9 knockout (KO) cells. Additionally, the level of beta 5i-containing intermediate proteasomes was significantly increased in these cells, as was beta 5i-related chymotrypsin-like activity. Moreover, proinsulin levels were restored in GRP94 KO upon beta 5i small interfering RNA-mediated knockdown. Finally, the fraction of beta-cells expressing the beta 5i-subunit is increased in human islets from type 2 diabetes patients. We conclude that beta 5i is an inducible proteasome subunit dedicated to the degradation of mishandled proinsulin.
KW - beta 5i
KW - GRP94
KW - proinsulin degradation
KW - proteasome
KW - restoration of proinsulin
KW - PROTEIN
U2 - 10.1152/ajpendo.00372.2019
DO - 10.1152/ajpendo.00372.2019
M3 - Journal article
C2 - 32255680
VL - 318
SP - E892-E900
JO - American Journal of Physiology - Endocrinology and Metabolism
JF - American Journal of Physiology - Endocrinology and Metabolism
SN - 0193-1849
IS - 6
ER -