%0 Journal Article %J Proc Natl Acad Sci U S A %D 2006 %T Genome-wide RNAi screen of Ca(2+) influx identifies genes that regulate Ca(2+) release-activated Ca(2+) channel activity. %A Zhang, Shenyuan L %A Yeromin, Andriy V %A Zhang, Xiang H-F %A Yu, Ying %A Safrina, Olga %A Penna, Aubin %A Roos, Jack %A Stauderman, Kenneth A %A Cahalan, Michael D %K Animals %K calcium %K Calcium Channels %K drosophila melanogaster %K Drosophila Proteins %K Enzyme Inhibitors %K Genome, Insect %K Humans %K Patch-Clamp Techniques %K Recombinant Fusion Proteins %K RNA Interference %K RNA, Double-Stranded %K Signal Transduction %K Thapsigargin %X

Recent studies by our group and others demonstrated a required and conserved role of Stim in store-operated Ca(2+) influx and Ca(2+) release-activated Ca(2+) (CRAC) channel activity. By using an unbiased genome-wide RNA interference screen in Drosophila S2 cells, we now identify 75 hits that strongly inhibited Ca(2+) influx upon store emptying by thapsigargin. Among these hits are 11 predicted transmembrane proteins, including Stim, and one, olf186-F, that upon RNA interference-mediated knockdown exhibited a profound reduction of thapsigargin-evoked Ca(2+) entry and CRAC current, and upon overexpression a 3-fold augmentation of CRAC current. CRAC currents were further increased to 8-fold higher than control and developed more rapidly when olf186-F was cotransfected with Stim. olf186-F is a member of a highly conserved family of four-transmembrane spanning proteins with homologs from Caenorhabditis elegans to human. The endoplasmic reticulum (ER) Ca(2+) pump sarco-/ER calcium ATPase (SERCA) and the single transmembrane-soluble N-ethylmaleimide-sensitive (NSF) attachment receptor (SNARE) protein Syntaxin5 also were required for CRAC channel activity, consistent with a signaling pathway in which Stim senses Ca(2+) depletion within the ER, translocates to the plasma membrane, and interacts with olf186-F to trigger CRAC channel activity.

%B Proc Natl Acad Sci U S A %V 103 %P 9357-62 %8 2006 Jun 13 %G eng %N 24 %1 http://www.ncbi.nlm.nih.gov/pubmed/16751269?dopt=Abstract %R 10.1073/pnas.0603161103