Details

Autor(en) / Beteiligte

• Zhang, S.
• Yeromin, A.
• Zhang, X. H.-F.
• Yu, Y.
• Safrina, O.
• Penna, A.
• Roos, J.
• Stauderman, K.
• Cahalan, M.

Titel

Genome-wide RNAi screen of Ca2+ influx identifies genes that regulate Ca2+ release-activated Ca2+ channel activity

Ist Teil von

• Proceedings of the National Academy of Sciences - PNAS, 2006-06, Vol.103 (24), p.9357-9362

Ort / Verlag

National Acad Sciences

Erscheinungsjahr

2006

Quelle

EZB Free E-Journals

Beschreibungen/Notizen

• 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. capacitative calcium entry (CCE) genome-wide screen CRAC channel RNA interference store-operated calcium (SOC) influx