Membrane stretching activates calcium permeability of a putative channel Pkd2 during fission yeast cytokinesis.

Pkd2 is the fission yeast homologue of polycystins. This putative ion channel localizes to the plasma membrane. It is required for the expansion of cell volume during interphase growth and cytokinesis, the last step of cell division. However, the channel activity of Pkd2 remains untested. Here, we examined the calcium permeability and mechanosensitivity of Pkd2 through in vitro reconstitution and calcium imaging of pkd2 mutant cells. Pkd2 was translated and inserted into the lipid bilayers of giant unilamellar vesicles using a cell-free expression system. The reconstituted Pkd2 permeated calcium when the membrane was stretched via hypoosmotic shock. In vivo, inactivation of Pkd2 through a temperature-sensitive mutation pkd2-B42 reduced the average intracellular calcium level by 34%. Compared with the wild type, the hypomorphic mutation pkd2-81KD reduced the amplitude of hypoosmotic shock-triggered calcium spikes by 59%. During cytokinesis, mutations of pkd2 reduced the calcium spikes, accompanying cell separation and the ensuing membrane stretching, by 60%. We concluded that fission yeast polycystin Pkd2 allows calcium influx when activated by membrane stretching, representing a likely mechanosensitive channel that contributes to the cytokinetic calcium spikes.

Cauliflower mosaic virus P6 inclusion body formation: A dynamic and intricate process.

During an infection, Cauliflower mosaic virus (CaMV) forms inclusion bodies (IBs) mainly composed of viral protein P6, where viral activities occur. Because viral processes occur in IBs, understanding the mechanisms by which they are formed is crucial. FL-P6 expressed in N. benthamiana leaves formed IBs of a variety of shapes and sizes. Small IBs were dynamic, undergoing fusion/dissociation events. Co-expression of actin-binding polypeptides with FL-P6 altered IB size distribution and inhibited movement. This suggests that IB movement is required for fusion and growth. A P6 deletion mutant was discovered that formed a single large IB per cell, which suggests it exhibited altered fusion/dissociation dynamics. Myosin-inhibiting drugs did not affect small IB movement, while those inhibiting actin polymerization did. Large IBs colocalized with components of the aggresome pathway, while small ones generally did not. This suggests a possible involvement of the aggresome pathway in large IB formation.

Phylogenetic patterns of foliar mineral nutrient accumulation among gypsophiles and their relatives in the Chihuahuan Desert.

PREMISE OF THE STUDY: Gypsum endemism in plants (gypsophily) is common on gypsum outcrops worldwide, but little is known about the functional ecology of Chihuahuan Desert gypsophiles. We investigated whether leaf chemistry of gypsophile lineages from the northern Chihuahuan Desert are similar to leaves of related nonendemic (gypsovag) species relative to their soil chemistry. We expected widely distributed gypsophiles (hypothesized to be older lineages on gypsum) would have distinct leaf chemistry from narrowly distributed, relatively younger lineages endemic to gypsum and gypsovags, reflecting adaptation to gypsum. METHODS: We collected leaves from 23 gypsophiles and related nonendemic taxa growing on nongypsum soils. Soils and leaves were analyzed for Ca, S, Mg, K, N, and P. Leaf gypsum was assessed using Fourier transform infrared spectroscopy. KEY RESULTS: Most widespread gypsophile lineages that are hypothesized to be relatively old accumulate foliar S, Ca, and gypsum, but younger gypsophile lineages and closely related gypsovags do not. Young, narrowly distributed gypsophile lineages have leaf chemical signatures similar to nonendemic congeners and confamilials. CONCLUSIONS: Our data suggest multiple adaptive mechanisms support life on gypsum in Chihuahuan Desert gypsophiles. Most widespread gypsophiles are specialized for life on gypsum, likely due to shared abilities to accumulate and assimilate S and Ca in leaves. In contrast, narrowly distributed gypsophiles may have mechanisms to exclude excess S and Ca from their leaves, preventing toxicity. Future work will investigate the nutrient accumulation and exclusion patterns of other plant organs to determine at what level excess S and Ca uptake is restricted for young-lineage gypsophiles and gypsovags.