Calpastatin is upregulated in non-immune neuronal cells via toll-like receptor 2 (TLR2) pathways by lipid-containing agonists

Calpain (intracellular Ca^2 +-dependent protease) and calpastatin (calpain specific endogenous inhibitor) are widely distributed in biological systems, and have been implicated in many cellular physiological and pathological processes. Calpastatin level is of central importance to the control of calpain activity. We demonstrated for the first time that calpastatin is overexpressed in mycoplasma-contaminated cultured cells (SH-SY5Y cells that are infected by a strain of Mycoplasma hyorhinis (NDMh)). We have found that the calpastatin-upregulating activity resides in the mycoplasmal membrane lipoproteins, and is associated with NF-κB activation. Calpain-promoted proteolysis is attenuated in the NDMh lipoprotein-treated cells. Here we show that the NDMh lipoproteins promoted an increase in calpastatin in SH-SY5Y cells via the TLR2/TAK1/NF-κB pathway. The synthetic mycoplasmal lipopeptide MALP-2 and the bacterial lipopeptide PAM3CSK4 (TLR2 agonists) also promoted calpastatin upregulation. LPS (TLR4 agonist) activated NF-κB without calpastatin increase in the cell. In contrast, lipoteichoic acid (TLR2 agonist) upregulated calpastatin not via NF-κB activation, but via the MEK1/ELK1 pathway. Zymosan and peptidoglycan, TLR2 agonists that lack lipids, did not induce calpastatin upregulation. Cell treatment with a calpastatin-upregulating agonist (lipoteichoic acid) led to the attenuation of Ca^2 +-promoted calpain activity, whereas agonists that do not upregulate calpastatin (LPS, Zymosan) were ineffective. Overall, the results indicate that in these non-immune cells, calpastatin is upregulated by TLR2-agonists containing lipids, with more than one downstream pathway involved. Such agonists may be useful for studying mechanisms and factors involved in calpastatin regulation. In addition, suitable TLR2 agonists may be of interest in devising treatments for pathological processes involving excessive calpain activation.     

Dominant plants alter the microclimate along a fog gradient in the Atacama Desert

Plant Ecology - We assessed the impact of fog on microclimate in a poorly understood fog desert under two common nurse plant species, at three sites: (1) foggy coast, (2) intermediate, and (3)...     

Saguaro (Carnegiea gigantea, Cactaceae) age-height relationships and growth: the development of a general growth curve

Because the growth rate of saguaros varies across different environments, past studies on saguaro population structure required extensive data collection (often over many decades) followed by site-specific analysis to estimate age at the sampled locale. However, when height-growth data from different populations are compared, the overall shape of the growth curves is similar. In this study, a formula was developed to establish saguaro age-height relationships (using stepwise regression) that can be applied to any saguaro population and only requires a site-specific factor to adjust the curve to the local growth rate. This adjustment factor can be established more efficiently and requires less data than the full analyses required for previous studies. Saguaro National Park East (SNP-E) was used as the baseline factor, set to 1.0. Results yielded a factor of 0.743 for SNP West. When the formula was applied to 10-yr interval data from Organ Pipe Cactus National Monument (OPCNM) in Arizona, USA, this location had a factor of 0.617 (relative to SNP-E). With this formula and relatively little field sampling, the age of any individual saguaro (whether the individual was sampled or not) in any population can be estimated.     

Normal mode analysis suggests a quaternary twist model for the nicotinic receptor gating mechanism

We present a three-dimensional model of the homopentameric alpha7 nicotinic acetylcholine receptor (nAChR), that includes the extracellular and membrane domains, developed by comparative modeling on the basis of: 1), the x-ray crystal structure of the snail acetylcholine binding protein, an homolog of the extracellular domain of nAChRs; and 2), cryo-electron microscopy data of the membrane domain collected on Torpedo marmorata nAChRs. We performed normal mode analysis on the complete three-dimensional model to explore protein flexibility. Among the first 10 lowest frequency modes, only the first mode produces a structural reorganization compatible with channel gating: a wide opening of the channel pore caused by a concerted symmetrical quaternary twist motion of the protein with opposing rotations of the upper (extracellular) and lower (transmembrane) domains. Still, significant reorganizations are observed within each subunit, that involve their bending at the domain interface, an increase of angle between the two beta-sheets composing the extracellular domain, the internal beta-sheet being significantly correlated to the movement of the M2 alpha-helical segment. This global symmetrical twist motion of the pentameric protein complex, which resembles the opening transition of other multimeric ion channels, reasonably accounts for the available experimental data and thus likely describes the nAChR gating process.     

Protein secondary structure assignment revisited: a detailed analysis of different assignment methods

Background  

A number of methods are now available to perform automatic assignment of periodic secondary structures from atomic coordinates, based on different characteristics of the secondary structures. In general these methods exhibit a broad consensus as to the location of most helix and strand core segments in protein structures. However the termini of the segments are often ill-defined and it is difficult to decide unambiguously which residues at the edge of the segments have to be included. In addition, there is a "twilight zone" where secondary structure segments depart significantly from the idealized models of Pauling and Corey. For these segments, one has to decide whether the observed structural variations are merely distorsions or whether they constitute a break in the secondary structure.     

Caspase-1 activity is required for neuronal differentiation of PC12 cells: cross-talk between the caspase and calpain systems

Previously, we have found that caspase-1 activity is increased during myoblast differentiation to myotubes. Here we show that caspase-1 activity is required for PC12 differentiation to neuronal-like cells. Caspase-1 is shown to be activated (by immunoblotting and by assessing activity in cell extracts) in the PC12 cells following the initial stage of differentiation. The inhibition of caspase-1 arrests PC12 cells at an intermediate stage of differentiation and prevents neurite outgrowth in these cells; the inhibition is reversed upon the removal of the inhibitor. Calpastatin (calpain endogenous specific inhibitor, and a known caspase substrate) is diminished at the later stages of PC12 cell differentiation, and diminution is prevented by caspase-1 inhibition. The degradation of fodrin (a known caspase and calpain substrate) is found in the advanced stage of differentiation. Caspase-1 has been implicated in the activation of proinflammatory cytokines, and in cell apoptosis. The involvement of caspase-1 in two distinct differentiation processes (myoblast fusion and neuronal differentiation of PC12 cells) indicates a function for this caspase in differentiation processes, and suggests some common mechanisms underlying caspase roles in such processes.     

Tightening of the ATP-binding sites induces the opening of P2X receptor channels

The opening of ligand-gated ion channels in response to agonist binding is a fundamental process in biology. In ATP-gated P2X receptors, little is known about the molecular events that couple ATP binding to channel opening. In this paper, we identify structural changes of the ATP site accompanying the P2X2 receptor activation by engineering extracellular zinc bridges at putative mobile regions as revealed by normal mode analysis. We provide evidence that tightening of the ATP sites shaped like open 'jaws' induces opening of the P2X ion channel. We show that ATP binding favours jaw tightening, whereas binding of a competitive antagonist prevents gating induced by this movement. Our data reveal the inherent dynamic of the binding jaw, and provide new structural insights into the mechanism of P2X receptor activation.     

Plant distribution and dispersal mechanisms at the Hassayampa River Preserve, Arizona, USA

This study focuses on the relationships between plant dispersal syndromes and plant distributions at the community scale. Species composition and cover are reported from 29 10 × 20-m vegetation plots along five topographic cross-sections in the riparian zone of the Hassayampa River Preserve, Arizona. We find that spatial patterns of dispersal guilds vary within the flood plain of this semiarid region river. Our main results are: (1) wind-dispersed species are fairly evenly distributed at all elevations and distances from the river, whereas cover of animal-dispersed species increases with elevation above, and at greater distances from, the river; (2) wind-dispersed species are proportionally more abundant in the pioneer Populus–Salix community, whereas plants in the late-seral Prosopis community are predominantly animal-dispersed; (3) most of the species classified as obligate-wetland and facultative-wetland are wind-dispersed, whereas facultative-upland and obligate-upland species are mostly animal-dispersed; and (4) there are significantly fewer wind-dispersed species in areas of high total vegetation cover. These results may reflect successional patterns resulting from periodic flooding. Low areas close to the river flood more frequently and with greater intensity than areas farther from the river. Many pioneer species that establish in disturbed areas are wind-dispersed. Over successional time, pioneer species cede to more drought tolerant species that are predominantly animal-dispersed.