STIM1L traps and gates Orai1 channels without remodeling the cortical ER

Journal of Cell Science
Sophie SaücNicolas Demaurex

Abstract

STIM proteins populate and expand cortical endoplasmic reticulum (ER) sheets to mediate store-operated Ca(2+) entry (SOCE) by trapping and gating Orai channels in ER-plasma membrane clusters. A longer splice variant, STIM1L, forms permanent ER-plasma membrane clusters and mediates rapid Ca(2+) influx in muscle. Here, we used electron microscopy, total internal reflection fluorescence (TIRF) microscopy and Ca(2+) imaging to establish the trafficking and signaling properties of the two STIM1 isoforms in Stim1(-/-)/Stim2(-/-) fibroblasts. Unlike STIM1, STIM1L was poorly recruited into ER-plasma membrane clusters and did not mediate store-dependent expansion of cortical ER cisternae. Removal of the STIM1 lysine-rich tail prevented store-dependent cluster enlargement, whereas inhibition of cytosolic Ca(2+) elevations or removal of the STIM1L actin-binding domain had no impact on cluster expansion. Finally, STIM1L restored robust but not accelerated SOCE and clustered with Orai1 channels more slowly than STIM1 following store depletion. These results indicate that STIM1L does not mediate rapid SOCE but can trap and gate Orai1 channels efficiently without remodeling cortical ER cisternae. The ability of STIM proteins to induce cortica...Continue Reading

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Citations

Jan 23, 2016·Frontiers in Pharmacology·Juan A RosadoIsaac Jardín
Apr 5, 2016·Cell Calcium·Rajesh BhardwajNicolas Demaurex
Aug 16, 2016·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Diana B PeckysNiels de Jonge
Oct 16, 2015·Experimental Biology and Medicine·Sarika Chaudhari, Rong Ma
Jan 31, 2016·American Journal of Physiology. Cell Physiology·Isabella DerlerChristoph Romanin
Feb 26, 2016·American Journal of Physiology. Cell Physiology·Barbara A Niemeyer
Nov 24, 2020·Current Opinion in Physiology·Marc FahrnerChristoph Romanin
Jan 6, 2021·International Journal of Molecular Sciences·Herwig GrabmayrMarc Fahrner
Feb 28, 2021·Pflügers Archiv : European journal of physiology·Paul RosenbergChaojian Wang
Dec 9, 2017·Advances in Biological Regulation·James W Putney
Oct 9, 2015·Journal of Cell Science·Daniele GuidoPaula Nunes

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Methods Mentioned

BETA
fluorescence imaging
electron microscopy
fluorescence-activated cell sorting
transfection
FCS
transfections

Software Mentioned

AnalySIS
Metamorph
GeneCust
NIS
Elements Ar software
ImageJ
Metafluor

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