STIM proteins populate and expand cortical endoplasmic reticulum (ER) sheets to mediate store‐operated Ca²⁺ entry (SOCE) by trapping and gating Orai channels in ER‐plasma membrane clusters. A longer splice variant, STIM1L, forms permanent ER‐plasma membrane clusters and mediates rapid Ca²⁺ influx in muscle. Here, we used electron microscopy, total internal reflection fluorescence (TIRF) microscopy and Ca²⁺ imaging to establish the trafficking and signaling properties of the two STIM1 isoforms in Stim1^−/−/Stim2^−/− fibroblasts. Unlike STIM1, STIM1L was poorly recruited into ER‐plasma membrane clusters and did not mediate store‐dependent expansion of cortical ER cisternae. Removal of the STIM1 lysine‐rich tail prevented store‐dependent cluster enlargement, whereas inhibition of cytosolic Ca²⁺ elevations or removal of the STIM1L actin‐binding domain had no impact on cluster expansion. Finally, STIM1L restored robust but not accelerated SOCE and clustered with Orai1 channels more slowly than STIM1 following store depletion. These results indicate that STIM1L does not mediate rapid SOCE but can trap and gate Orai1 channels efficiently without remodeling cortical ER cisternae. The ability of STIM proteins to induce cortical ER formation is dispensable for SOCE and requires the lysine‐rich tail of STIM1 involved in binding to phosphoinositides.

To initiate studies on how protein-protein interaction (or “interactome”) networks relate to multicellular functions, we have mapped a large fraction of the Caenorhabditis elegans interactome network. Starting with a subset of metazoan-specific proteins, more than 4000 interactions were identified from high-throughput, yeast two-hybrid (HT=Y2H) screens. Independent coaffinity purification assays experimentally validated the overall quality of this Y2H data set. Together with already described Y2H interactions and interologs predicted in silico , the current version of the Worm Interactome (WI5) map contains ∼5500 interactions. Topological and biological features of this interactome network, as well as its integration with phenome and transcriptome data sets, lead to numerous biological hypotheses.

Current yeast interactome network maps contain several hundred molecular complexes with limited and somewhat controversial representation of direct binary interactions. We carried out a comparative quality assessment of current yeast interactome data sets, demonstrating that high-throughput yeast two-hybrid (Y2H) screening provides high-quality binary interaction information. Because a large fraction of the yeast binary interactome remains to be mapped, we developed an empirically controlled mapping framework to produce a “second-generation” high-quality, high-throughput Y2H data set covering ∼20% of all yeast binary interactions. Both Y2H and affinity purification followed by mass spectrometry (AP/MS) data are of equally high quality but of a fundamentally different and complementary nature, resulting in networks with different topological and biological properties. Compared to co-complex interactome models, this binary map is enriched for transient signaling interactions and intercomplex connections with a highly significant clustering between essential proteins. Rather than correlating with essentiality, protein connectivity correlates with genetic pleiotropy.