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 -