The inducible beta 5i proteasome subunit contributes to proinsulin degradation in GRP94-deficient beta-cells and is overexpressed in type 2 diabetes pancreatic islets

Muhammad Saad Khilji, Sophie Emilie Bresson, Danielle Verstappen, Celina Pihl, Phillip Alexander Keller Andersen, Jette Bach Agergaard, Tina Dahlby, Tenna Holgersen Bryde, Kristian Klindt, Christian Kronborg Nielsen, Anna Walentinsson, Dusan Zivkovic, Marie-Pierre Bousquet, Bjorn Tyrberg, Sarah J. Richardson, Noel G. Morgan, Thomas Mandrup-Poulsen, Michal Tomasz Marzec*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

8 Citations (Scopus)

Abstract

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.

Original languageEnglish
JournalAmerican Journal of Physiology: Endocrinology and Metabolism
Volume318
Issue number6
Pages (from-to)E892-E900
ISSN0193-1849
DOIs
Publication statusPublished - 2020

Keywords

  • beta 5i
  • GRP94
  • proinsulin degradation
  • proteasome
  • restoration of proinsulin
  • PROTEIN

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