Genome-wide RNAi screen of Ca(2+) influx identifies genes that regulate Ca(2+) release-activated Ca(2+) channel activity. | Academic Article individual record
abstract

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.

author list (cited authors)
Zhang, S. L., Yeromin, A. V., Zhang, X., Yu, Y., Safrina, O., Penna, A., ... Cahalan, M. D.
publication date
2006
keywords
  • Enzyme Inhibitors
  • Calcium
  • Genome, Insect
  • RNA Interference
  • Thapsigargin
  • Humans
  • Drosophila Proteins
  • Recombinant Fusion Proteins
  • RNA, Double-Stranded
  • Signal Transduction
  • Patch-Clamp Techniques
  • Drosophila Melanogaster
  • Animals
  • Calcium Channels
altmetric score

9.0

citation count

658