The NH2 terminus regulates voltage-dependent gating of CALHM ion channels

Calcium homeostasis modulator protein-1 (CALHM1) and its Caenorhabditis elegans (ce) homolog, CLHM-1, belong to a new family of physiologically important ion channels that are regulated by voltage and extracellular Ca2+ (Ca2+o) but lack a canonical voltage-sensing domain. Consequently, the intrinsic voltage-dependent gating mechanisms for CALHM channels are unknown. Here, we performed voltage-clamp experiments on ceCLHM-1 chimeric, deletion, insertion, and point mutants to assess the role of the NH2 terminus (NT) in CALHM channel gating. Analyses of chimeric channels in which the ceCLHM-1 and human (h)CALHM1 NH2 termini were interchanged showed that the hCALHM1 NT destabilized channel-closed states, whereas the ceCLHM-1 NT had a stabilizing effect. In the absence of Ca2+o, deletion of up to eight amino acids from the ceCLHM-1 NT caused a hyperpolarizing shift in the conductance-voltage relationship with little effect on voltage-dependent slope. However, deletion of nine or more amino acids decreased voltage dependence and induced a residual conductance at hyperpolarized voltages. Insertion of amino acids into the NH2-terminal helix also decreased voltage dependence but did not prevent channel closure. Mutation of ceCLHM-1 valine 9 and glutamine 13 altered half-maximal activation and voltage dependence, respectively, in 0 Ca2+. In 2 mM Ca2+o, ceCLHM-1 NH2-terminal deletion and point mutant channels closed completely at hyperpolarized voltages with apparent affinity for Ca2+o indistinguishable from wild-type ceCLHM-1, although the ceCLHM-1 valine 9 mutant exhibited an altered conductance-voltage relationship and kinetics. We conclude that the NT plays critical roles modulating voltage dependence and stabilizing the closed states of CALHM channels.

Short-term Flooding Affects Gas Exchange Characteristics of Containerized Sour Cherry Trees

The effects of short-term soil flooding on gas exchange characteristics of containerized sour cherry trees (Prunus cerasus L. cv. Montmorency /P. mahaleb L.) were studied under laboratory conditions. Soil flooding reduced net CO2 assimilation (A) within 24 hours. Net CO2 assimilation and residual conductance to CO2(gr′) declined to ≈30% of control values after 5 days of flooding. Effects on stomatal conductance to CO2 (gS) and intercellular CO2 (Ci) were not significant during the 5 days of treatment. Apparent quantum yield (Φ) gradually declined to 52% that of controls during these 5 days. In a second experiment, CO2 response curves suggested that, initially, stomatal and nonstomatal limitations to A were of about equal importance; however, as flooding continued, nonstomatal limitations became dominant.

Gas Exchange Characteristics of Fragaria chiloensis Genotypes

Twenty-five female clones of Fragaria chiloensis (L.) Duchesene collected from the California and Oregon coasts were surveyed for gas exchange rates under field conditions. Carbon assimilation (A) rates of native clones were 25% to 69% higher than for `Totem' (Fragaria × ananassa Duchesne) on a leaf-area basis (μmol of CO2 per sec/m2) and 7% to 77% higher when expressed on a leaf dry-weight basis (μmol of CO2 per kg dry wt/sec). Higher rates of stomatal conductance (gs) were observed in 16 of 25 F. chiloensis clones than in `Totem', with nine of 25 clones also having higher rates of transpiration (E). All clones had higher rates of residual conductance (gr) and greater water use efficiency (WUE) than the cultivated standard. The gas exchange characteristics of four strawberry cultivars (F. × ananassa) and four F. chiloensis genotypes were compared under standard greenhouse conditions. F. chiloensis genotypes had higher rates of A than cultivars when expressed on per leaf-area and dry-weight bases. Native clones also had higher rates of gs, gr, E, and WUE and greater quantum yield. Differences in chlorophyll content were observed among genotypes, but not between species.