Sarco(endo)plasmic reticulum ATPase is a molecular partner of Wolfram syndrome 1 protein, which negatively regulates its expression

Malgorzata Zatyka, G Da Silva Xavier, Elisa A Bellomo, Wendy Leadbeater, Dewi Astuti, Joel Smith, Francesco Michelangeli, Guy A Rutter, Timothy Barrett

Research output: Contribution to journalArticlepeer-review

24 Citations (Scopus)
156 Downloads (Pure)

Abstract

Wolfram syndrome is an autosomal recessive disorder characterized by neurodegeneration and diabetes mellitus. The gene responsible for the syndrome (WFS1) encodes an endoplasmic reticulum (ER)-resident transmembrane protein that is involved in the regulation of the unfolded protein response (UPR), intracellular ion homeostasis, cyclic adenosine monophosphate production and regulation of insulin biosynthesis and secretion. In this study, single cell Ca(2+) imaging with fura-2 and direct measurements of free cytosolic ATP concentration ([ATP]CYT) with adenovirally expressed luciferase confirmed a reduced and delayed rise in cytosolic free Ca(2+) concentration ([Ca(2+)]CYT), and additionally, diminished [ATP]CYT rises in response to elevated glucose concentrations in WFS1-depleted MIN6 cells. We also observed that sarco(endo)plasmic reticulum ATPase (SERCA) expression was elevated in several WFS1-depleted cell models and primary islets. We demonstrated a novel interaction between WFS1 and SERCA by co-immunoprecipitation in Cos7 cells and with endogenous proteins in human neuroblastoma cells. This interaction was reduced when cells were treated with the ER stress inducer dithiothreitol. Treatment of WFS1-depleted neuroblastoma cells with the proteasome inhibitor MG132 resulted in reduced accumulation of SERCA levels compared with wild-type cells. Together these results reveal a role for WFS1 in the negative regulation of SERCA and provide further insights into the function of WFS1 in calcium homeostasis.

Original languageEnglish
Pages (from-to)814-827
Number of pages14
JournalHuman Molecular Genetics
Volume24
Issue number3
Early online date30 Sept 2014
DOIs
Publication statusPublished - 1 Feb 2015

Keywords

  • Animals
  • COS Cells
  • Calcium/metabolism
  • Cell Line, Tumor
  • Cells, Cultured
  • Cercopithecus aethiops
  • Dithiothreitol/pharmacology
  • Gene Expression Regulation
  • Humans
  • Insulin/secretion
  • Membrane Proteins/metabolism
  • Mice
  • Mice, Knockout
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases/metabolism

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