Engineered mutations change the structure and stability of a virus-like particle

The single-coat protein (CP) of bacteriophage Qβ self-assembles into T = 3 icosahedral virus-like particles (VLPs), of interest for a wide range of applications. These VLPs are very stable, but identification of the specific molecular determinants of this stability is lacking. To investigate these determinants along with manipulations that confer more capabilities to our VLP material, we manipulated the CP primary structure to test the importance of various putative stabilizing interactions. Optimization of a procedure to incorporate fused CP subunits allowed for good control over the average number of covalent dimers in each VLP. We confirmed that the disulfide linkages are the most important stabilizing elements for the capsid and that acidic conditions significantly enhance the resistance of VLPs to thermal degradation. Interdimer interactions were found to be less important for VLP assembly than intradimer interactions. Finally, a single point mutation in the CP resulted in a population of smaller VLPs in three distinct structural forms.     

Leakage induced in Escherichia coli cells by A protein-RNA complexes from bacteriophage f2

Complexes of f2 phage RNA and its A protein, or maturation protein, transfect Escherichia coli cells much better than does protein-free RNA. We used these complexes to introduce the bacteriophage f2 lysis gene into cells. The A protein-RNA complex was found to kill cells, probably by causing them to leak large macromolecules. Previously induced beta-galactosidase leaked from cells treated either with the A protein-RNA complex or with lethal but noninfectious complexes that had been treated with formaldehyde. This observation was consistent with an earlier finding that formaldehyde-treated f2 RNA stimulates the in vitro synthesis of a lysis protein. The complexes did not stimulate the rate of leakage of beta-galactosidase from a streptomycin-resistant mutant known to be lysis defective. On the other hand, the rate of leakage was increased in a double mutant resistant to both streptomycin and rifampin and which is lysed normally by f2 bacteriophage.     

Personality and the emergence of the pace-of-life syndrome concept at the population level

The pace-of-life syndrome (POLS) hypothesis specifies that closely related species or populations experiencing different ecological conditions should differ in a suite of metabolic, hormonal and immunity traits that have coevolved with the life-history particularities related to these conditions. Surprisingly, two important dimensions of the POLS concept have been neglected: (i) despite increasing evidence for numerous connections between behavioural, physiological and life-history traits, behaviours have rarely been considered in the POLS yet; (ii) the POLS could easily be applied to the study of covariation among traits between individuals within a population. In this paper, we propose that consistent behavioural differences among individuals, or personality, covary with life history and physiological differences at the within-population, interpopulation and interspecific levels. We discuss how the POLS provides a heuristic framework in which personality studies can be integrated to address how variation in personality traits is maintained within populations.     

Does climate control the northern range limit of eastern white cedar (Thuja occidentalis L.)?

Our aim was to test whether or not climate influence the northern distributional limit of eastern white cedar (Thuja occidentalis L.) by affecting its radial growth and regeneration. Twenty-two sites were selected along the bioclimatic gradient in Northwestern Québec, Canada. The gradient was divided into three zones based on the abundance of white cedar stands: continuous, discontinuous, and marginal zones. Radial growth and regeneration (numbers of sedds, saplings) were determined for each zone. Results showed that basal area increment (b.a.i.) was the same along the gradient. Seed production and germination did not differ between zones. One-year-old seedlings and small individuals (<30 cm) were less abundant in the northern sites than in stands from the continuous and discontinuous zones. More saplings were found in the northernmost sites (389/400 m² in the marginal and 354.7/400 m² in the discontinuous zones) than in the south (130.6/400 m²). Layering seemed to compensate for the low recruitment observed in the marginal sites. Recruitment of seedlings originating from sexual reproduction in the discontinuous zone was not different from the stands in the southern areas. Thus, a climatically driven decrease in recruitment cannot explain the observed decrease in white cedar abundance, which occurred at this latitude. Although seedlings were less abundant in isolated northern marginal stands, sexually based regeneration was still possible. Therefore, the direct effect of climate seems to only have a minor influence on white cedar northern distributional limit and other factors, such as natural disturbances, might better explain its actual boundary.     

An ERP Study of the Processing of Common and Decimal Fractions: How Different They Are

This study explored event-related potential (ERP) correlates of common fractions (1/5) and decimal fractions (0.2). Thirteen subjects performed a numerical magnitude matching task under two conditions. In the common fraction condition, a nonsymbolic fraction was asked to be judged whether its magnitude matched the magnitude of a common fraction; in the decimal fraction condition, a nonsymbolic fraction was asked to be matched with a decimal fraction. Behavioral results showed significant main effects of condition and numerical distance, but no significant interaction of condition and numerical distance. Electrophysiological data showed that when nonsymbolic fractions were compared to common fractions, they displayed larger N1 and P3 amplitudes than when they were compared to decimal fractions. This finding suggested that the visual identification for nonsymbolic fractions was different under the two conditions, which was not due to perceptual differences but to task demands. For symbolic fractions, the condition effect was observed in the N1 and P3 components, revealing stimulus-specific visual identification processing. The effect of numerical distance as an index of numerical magnitude representation was observed in the P2, N3 and P3 components under the two conditions. However, the topography of the distance effect was different under the two conditions, suggesting stimulus specific semantic processing of common fractions and decimal fractions.     

Intra-Genomic Variation in the Ribosomal Repeats of Nematodes

Ribosomal loci represent a major tool for investigating environmental diversity and community structure via high-throughput marker gene studies of eukaryotes (e.g. 18S rRNA). Since the estimation of species’ abundance is a major goal of environmental studies (by counting numbers of sequences), understanding the patterns of rRNA copy number across species will be critical for informing such high-throughput approaches. Such knowledge is critical, given that ribosomal RNA genes exist within multi-copy repeated arrays in a genome. Here we measured the repeat copy number for six nematode species by mapping the sequences from whole genome shotgun libraries against reference sequences for their rRNA repeat. This revealed a 6-fold variation in repeat copy number amongst taxa investigated, with levels of intragenomic variation ranging from 56 to 323 copies of the rRNA array. By applying the same approach to four C. elegans mutation accumulation lines propagated by repeated bottlenecking for an average of ~400 generations, we find on average a 2-fold increase in repeat copy number (rate of increase in rRNA estimated at 0.0285-0.3414 copies per generation), suggesting that rRNA repeat copy number is subject to selection. Within each Caenorhabditis species, the majority of intragenomic variation found across the rRNA repeat was observed within gene regions (18S, 28S, 5.8S), suggesting that such intragenomic variation is not a product of selection for rRNA coding function. We find that the dramatic variation in repeat copy number among these six nematode genomes would limit the use of rRNA in estimates of organismal abundance. In addition, the unique pattern of variation within a single genome was uncorrelated with patterns of divergence between species, reflecting a strong signature of natural selection for rRNA function. A better understanding of the factors that control or affect copy number in these arrays, as well as their rates and patterns of evolution, will be critical for informing estimates of global biodiversity.     

Synthesis, maturation, and trafficking of human Na+-dicarboxylate cotransporter NaDC1 requires the chaperone activity of cyclophilin B

Renal excretion of citrate, an inhibitor of calcium stone formation, is controlled mainly by reabsorption via the apical Na⁺-dicarboxylate cotransporter NaDC1 (SLC13A2) in the proximal tubule. Recently, it has been shown that the protein phosphatase calcineurin inhibitors cyclosporin A (CsA) and FK-506 induce hypocitraturia, a risk factor for nephrolithiasis in kidney transplant patients, but apparently through urine acidification. This suggests that these agents up-regulate NaDC1 activity. Using the Xenopus lævis oocyte and HEK293 cell expression systems, we examined first the effect of both anti-calcineurins on NaDC1 activity and expression. While FK-506 had no effect, CsA reduced NaDC1-mediated citrate transport by lowering heterologous carrier expression (as well as endogenous carrier expression in HEK293 cells), indicating that calcineurin is not involved. Given that CsA also binds specifically to cyclophilins, we determined next whether such proteins could account for the observed changes by examining the effect of selected cyclophilin wild types and mutants on NaDC1 activity and cyclophilin-specific siRNA. Interestingly, our data show that the cyclophilin isoform B is likely responsible for down-regulation of carrier expression by CsA and that it does so via its chaperone activity on NaDC1 (by direct interaction) rather than its rotamase activity. We have thus identified for the first time a regulatory partner for NaDC1, and have gained novel mechanistic insight into the effect of CsA on renal citrate transport and kidney stone disease, as well as into the regulation of membrane transporters in general.