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

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

*Corresponding author for this work

Research output: Contribution to journalArticle

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 Ca2+ 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 Ca2+ concentration ([Ca2+]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
Article numberddu499
Pages (from-to)814-827
Number of pages14
JournalHuman Molecular Genetics
Volume24
Issue number3
DOIs
Publication statusPublished - 30 Sep 2014

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    Zatyka, M., Da Silva Xavier, G., Bellomo, E. A., Leadbeater, W., Astuti, D., Smith, J., Michelangeli, F., Rutter, G. A., & Barrett, T. G. (2014). Sarco(endo)plasmic reticulum atpase is a molecular partner of wolfram syndrome 1 protein, which negatively regulates its expression. Human Molecular Genetics, 24(3), 814-827. [ddu499]. https://doi.org/10.1093/hmg/ddu499