SUMMARY Most available information on ER-plasma membrane (PM) contacts in cells of higher eukaryotes concerns proteins implicated in the regulation of Ca2+ entry. However, growing evidence suggests that such contacts play more general roles in cell physiology, pointing to the existence of additionally ubiquitously expressed ER-PM tethers. Here we show that the three Extended-Synaptotagmins (E-Syts) are ER proteins that participate in such tethering function via C2 domain-dependent interactions with the PM that require PI(4,5)P2 in the case of E-Syt2 and E-Syt3 and also elevation of cytosolic Ca2+ in the case of E-Syt1. As they form heteromeric complexes, the E-Syts confer cytosolic Ca2+ regulation to ER-PM contact formation. E-Syts-dependent contacts, however, are not required for store-operated Ca2+ entry. Thus, the ER-PM tethering function of the E-Syts (tricalbins in yeast), mediate the formation of ER-PM contacts sites which are functionally distinct from those mediated by STIM1 and Orai1.

Multiple modalities converge on a common gate to control K2P channel functionK2P potassium channels play important roles in the regulation of neuronal excitability. K2P channels are gated chemical, thermal, and mechanical stimuli, and the present study identifies and characterizes a common molecular gate that responds to all different stimuli, both activating and inhibitory ones.

Metabolic and thermal stimuli control K2P2.1 (TREK-1) through modular sensory and gating domainsThe two-pore domain potassium channel K2P2.1 (TREK-1) responds to extracellular and intracellular stimuli, including pH and temperature. This study elucidates how the intracellular sensor element relays metabolic and thermal stimuli to the extracellular C-type gating element.

No abstract

The [PSI(+)] prion is the aggregated self-propagating form of the Sup35 protein from the yeast Saccharomyces cerevisiae. Aggregates of Sup35 in [PSI(+)] cells exist in different heritable conformations, called "variants," and they are composed of detergent-resistant Sup35 polymers, which may be closely associated with themselves, other proteins, or both. Here, we report that disassembly of the aggregates into individual Sup35 polymers and non-Sup35 components increases their infectivity while retaining their variant specificity, showing that variant-specific [PSI(+)] infection can be transmitted by Sup35 polymers alone. Morphological analysis revealed that Sup35 isolated from [PSI(+)] yeast has the appearance of short barrels, and bundles, which seem to be composed of barrels. We show that the major components of two different variants of [PSI(+)] are interacting infectious Sup35 polymers and Ssa1/2. Using a candidate approach, we detected Hsp104, Ssb1/2, Sis1, Sse1, Ydj1, and Sla2 among minor components of the aggregates. We demonstrate that Ssa1/2 efficiently binds to the prion domain of Sup35 in [PSI(+)] cells, but that it interacts poorly with the nonaggregated Sup35 found in [psi(-)] cells. Hsp104, Sis1, and Sse1 interact preferentially with the prion versus nonprion form of Sup35, whereas Sla2 and Ssb1/2 interact with both forms of Sup35 with similar efficiency.