(679.12) S-acylation of STIM1 regulates store-operated calcium entry

Poster Board Number: A473

Goutham Kodakandla (Cooper Medical School of Rowan University), Savannah West (University of Texas Health Science Center at Houston), Qioachu Wang (University of Texas Health Science Center at Houston), Ritika Tewari (University of Texas Health Science Center at Houston), Michael Zhu (University of Texas Health Science Center at Houston), Askar Akimzhanov (University of Texas Health Science Center at Houston), Darren Boehning (Cooper Medical School of Rowan University)

The initiating events happening during store-operated calcium entry are not completely
understood despite the tremendous importance of this process in immune responses. Upon
endoplasmic reticulum (ER) calcium depletion, STIM1 and Orai1 form calcium release-activated calcium (CRAC) channels to allow calcium influx from the extracellular milieu. S-acylation is a reversible post-translational modification of cysteine residues that affects protein structure, stability, and targeting to subdomains in the plasma membrane. Previously, we have shown that Orai1 is rapidly S-acylated following T cell activation in Jurkat cells as well as ER calcium store depletion in HEK293 cells. Here we demonstrate that STIM1 is S-acylated at C437 residue upon T cell activation and ER calcium store depletion. This is an interesting observation because very little is known about S-acylation and its consequences in ER-resident proteins. Upon store depletion, STIM1 adopts an extended conformation that permits binding to Orai1. We hypothesize that S-acylation of STIM1 promotes and stabilizes the extended conformation of STIM1. Consistent with this hypothesis, we show that substitution of cysteine 437 in the STIM1 Orai1 activating region to a serine decreases calcium entry after store depletion. We further show that the ability of STIM1 to form puncta depends on its S-acylation at this residue. Together with S-acylation of Orai1, these data demonstrate that CRAC channel components are rapidly S-acylated after ER calcium store depletion. In summary, based on our previous published literature on Orai1 S-acylation, and current discovery of S-acylation of STIM1, it is evident that S-acylation recruits Orai1 to specialized membrane subdomains to form puncta with STIM1, whose extended conformation is stabilized by S-acylation at its 437 residue. Hence, S-acylation of Orai1 and STIM1 play a crucial role facilitate store-operated calcium entry.

Support or Funding Information

NIH/NIGMS - 5R01GM130840

lt;pgt;NIH/NIGMS - 5R01GM130840lt;/pgt;