Nanoscale patterning of STIM1 and Orai1 during store-operated Ca2+ entry

Proceedings of the National Academy of Sciences of the United States of America
Stefano PerniC Franzini-Armstrong

Abstract

Stromal interacting molecule (STIM) and Orai proteins constitute the core machinery of store-operated calcium entry. We used transmission and freeze-fracture electron microscopy to visualize STIM1 and Orai1 at endoplasmic reticulum (ER)-plasma membrane (PM) junctions in HEK 293 cells. Compared with control cells, thin sections of STIM1-transfected cells possessed far more ER elements, which took the form of complex stackable cisternae and labyrinthine structures adjoining the PM at junctional couplings (JCs). JC formation required STIM1 expression but not store depletion, induced here by thapsigargin (TG). Extended molecules, indicative of STIM1, decorated the cytoplasmic surface of ER, bridged a 12-nm ER-PM gap, and showed clear rearrangement into small clusters following TG treatment. Freeze-fracture replicas of the PM of Orai1-transfected cells showed extensive domains packed with characteristic "particles"; TG treatment led to aggregation of these particles into sharply delimited "puncta" positioned upon raised membrane subdomains. The size and spacing of Orai1 channels were consistent with the Orai crystal structure, and stoichiometry was unchanged by store depletion, coexpression with STIM1, or an Orai1 mutation (L273D) a...Continue Reading

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Citations

Jan 15, 2017·Cell Calcium·Yandong ZhouDonald L Gill
Mar 7, 2017·Cell Calcium·Woo Young ChungShmuel Muallem
May 31, 2017·Biochimica Et Biophysica Acta. Molecular Cell Research·Chi-Lun ChangJen Liou
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Oct 17, 2017·EMBO Reports·Shmuel MuallemMalini Ahuja
Oct 2, 2019·Cold Spring Harbor Perspectives in Biology·Richard S Lewis
Oct 27, 2016·The Journal of Biological Chemistry·Xiangyu CaiDonald L Gill
Mar 28, 2018·Proceedings of the National Academy of Sciences of the United States of America·Yandong ZhouDonald L Gill
Oct 29, 2019·ELife·Antonio MichelucciFeliciano Protasi
Aug 20, 2020·Journal of Muscle Research and Cell Motility·Feliciano ProtasiSimona Boncompagni
Jun 27, 2020·The Journal of General Physiology·Joseph L DynesMichael D Cahalan
Feb 22, 2017·Scientific Reports·Pascal YazbeckDolly Mehta
Oct 20, 2020·Neuropathology : Official Journal of the Japanese Society of Neuropathology·Georges Arielle PecheJohann Böhm
Sep 22, 2020·Current Opinion in Physiology·James H BaraniakDonald L Gill
Jan 21, 2021·International Journal of Molecular Sciences·Diana B PeckysNiels de Jonge
Mar 3, 2021·Proceedings of the National Academy of Sciences of the United States of America·Yihan ShenColin W Taylor
Jul 3, 2021·International Journal of Molecular Sciences·Feliciano ProtasiSimona Boncompagni

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