The VgrG Proteins Are “à la Carte” Delivery Systems for Bacterial Type VI Effectors

The bacterial type VI secretion system (T6SS) is a supra-molecular complex akin to bacteriophage tails, with VgrG proteins acting as a puncturing device. The Pseudomonas aeruginosa H1-T6SS has been extensively characterized. It is involved in bacterial killing and in the delivery of three toxins, Tse1–3. Here, we demonstrate the independent contribution of the three H1-T6SS co-regulated vgrG genes, vgrG1abc, to bacterial killing. A putative toxin is encoded in the vicinity of each vgrG gene, supporting the concept of specific VgrG/toxin couples. In this respect, VgrG1c is involved in the delivery of an Rhs protein, RhsP1. The RhsP1 C terminus carries a toxic activity, from which the producing bacterium is protected by a cognate immunity. Similarly, VgrG1a-dependent toxicity is associated with the PA0093 gene encoding a two-domain protein with a putative toxin domain (Toxin_61) at the C terminus. Finally, VgrG1b-dependent killing is detectable upon complementation of a triple vgrG1abc mutant. The VgrG1b-dependent killing is mediated by PA0099, which presents the characteristics of the superfamily nuclease 2 toxin members. Overall, these data develop the concept that VgrGs are indispensable components for the specific delivery of effectors. Several additional vgrG genes are encoded on the P. aeruginosa genome and are not linked genetically to other T6SS genes. A closer inspection of these clusters reveals that they also encode putative toxins. Overall, these associations further support the notion of an original form of secretion system, in which VgrG acts as the carrier.     

More Productive Than Maize in the Midwest: How Does Miscanthus Do It?

In the first side-by-side large-scale trials of these two C₄ crops in the U.S. Corn Belt, Miscanthus (Miscanthus × giganteus) was 59% more productive than grain maize (Zea mays). Total productivity is the product of the total solar radiation incident per unit land area and the efficiencies of light interception (ε_i) and its conversion into aboveground biomass (ε_ca). Averaged over two growing seasons, ε_ca did not differ, but ε_i was 61% higher for Miscanthus, which developed a leaf canopy earlier and maintained it later. The diurnal course of photosynthesis was measured on sunlit and shaded leaves of each species on 26 dates. The daily integral of leaf-level photosynthetic CO₂ uptake differed slightly when integrated across two growing seasons but was up to 60% higher in maize in mid-summer. The average leaf area of Miscanthus was double that of maize, with the result that calculated canopy photosynthesis was 44% higher in Miscanthus, corresponding closely to the biomass differences. To determine the basis of differences in mid-season leaf photosynthesis, light and CO₂ responses were analyzed to determine in vivo biochemical limitations. Maize had a higher maximum velocity of phosphoenolpyruvate carboxylation, velocity of phosphoenolpyruvate regeneration, light saturated rate of photosynthesis, and higher maximum quantum efficiency of CO₂ assimilation. These biochemical differences, however, were more than offset by the larger leaf area and its longer duration in Miscanthus. The results indicate that the full potential of C₄ photosynthetic productivity is not achieved by modern temperate maize cultivars.     

Exosome Secretion: The Art of Reutilizing Nonrecycled Proteins?

Multivesicular bodies contain membrane vesicles which either undergo lysosomal digestion or are released in the extracellular environment as exosomes. Evidence is accumulating that supports a physiological role for exosomes in, for example, antigen presentation or removal of transferrin receptor during reticulocyte development. Here, inspired by observations on exosomal release from reticulocytes, we discuss the potential involvement of the so-called ESCRT mechanism in the entrapment of both lysosomal and exosomal cargo within the intralumenal vesicles of multivesicular bodies. We propose that this mechanism operates at different sites in the endocytic itinerary in different cells, thereby providing a tool for directional sorting. We also explore the possibility that the efficiency of sorting of molecules into exosomes increases when the recycling kinetics of molecules decreases, exosomal sorting being favored by intermolecular interactions occurring within lipid domains, or with protein webs, that slow lateral mobility. These considerations are mirrored in the context of current knowledge on the mechanism of protein sorting for degradation in lysosomes, and the hijacking of such mechanisms by some retroviruses for particle budding.     

The 2.5 ? Structure of the Enterococcus Conjugation Protein TraM resembles VirB8 Type IV Secretion Proteins

Conjugative plasmid transfer is the most important means of spreading antibiotic resistance and virulence genes among bacteria and therefore presents a serious threat to human health. The process requires direct cell-cell contact made possible by a multiprotein complex that spans cellular membranes and serves as a channel for macromolecular secretion. Thus far, well studied conjugative type IV secretion systems (T4SS) are of Gram-negative (G−) origin. Although many medically relevant pathogens (e.g., enterococci, staphylococci, and streptococci) are Gram-positive (G+), their conjugation systems have received little attention. This study provides structural information for the transfer protein TraM of the G+ broad host range Enterococcus conjugative plasmid pIP501. Immunolocalization demonstrated that the protein localizes to the cell wall. We then used opsonophagocytosis as a novel tool to verify that TraM was exposed on the cell surface. In these assays, antibodies generated to TraM recruited macrophages and enabled killing of pIP501 harboring Enteroccocus faecalis cells. The crystal structure of the C-terminal, surface-exposed domain of TraM was determined to 2.5 Å resolution. The structure, molecular dynamics, and cross-linking studies indicated that a TraM trimer acts as the biological unit. Despite the absence of sequence-based similarity, TraM unexpectedly displayed a fold similar to the T4SS VirB8 proteins from Agrobacterium tumefaciens and Brucella suis (G−) and to the transfer protein TcpC from Clostridium perfringens plasmid pCW3 (G+). Based on the alignments of secondary structure elements of VirB8-like proteins from mobile genetic elements and chromosomally encoded T4SS from G+ and G− bacteria, we propose a new classification scheme of VirB8-like proteins.     

Building Ribosomes: Even More Expensive Than Expected?

Recent quantitative analyses of ribosomal protein trafficking in HeLa cells have revealed a prominent and unexpected role for the proteasome in regulating the availability of ribosomal proteins for subunit assembly.     

Do Unique Proteins Exist in Taste Buds?

Proteins in papillae on the bovine tongue were analyzed by semi-micro, polyacrylamide gel electrophoresis. All the proteins in the papillae with taste buds were observed to be common to proteins in the surrounding epithelium without taste buds. The protein band which was reported to form a weak complex with compounds called sweet by man was also found in all parts of the tongue epithelium. The receptor molecules for chemical stimuli may be distributed in all the cells of the tongue epithelium or the content of receptor molecules in taste bud papillae may be extremely low.     

Why Are Some Plant Genera More Invasive Than Others?

Determining how biological traits are related to the ability of groups of organisms to become economically damaging when established outside of their native ranges is a major goal of population biology, and important in the management of invasive species. Little is known about why some taxonomic groups are more likely to become pests than others among plants. We investigated traits that discriminate vascular plant genera, a level of taxonomic generality at which risk assessment and screening could be more effectively performed, according to the proportion of naturalized species which are pests. We focused on the United States and Canada, and, because our purpose is ultimately regulatory, considered species classified as weeds or noxious. Using contingency tables, we identified 11 genera of vascular plants that are disproportionately represented by invasive species. Results from boosted regression tree analyses show that these categories reflect biological differences. In summary, approximately 25% of variation in genus proportions of weeds or noxious species was explained by biological covariates. Key explanatory traits included genus means for wetland habitat affinity, chromosome number, and seed mass.     

Do More Active Children Sleep More? A Repeated Cross-Sectional Analysis Using Accelerometry

Aim

To determine whether levels of daytime physical activity are associated with sleep duration and night waking in children assessed using accelerometry, and if these associations change over time.

Methods

24-hour accelerometry data were obtained from 234 children at 3, 5 and 7 years of age for at least 5 days at each time. Sleep duration was estimated using the Sadeh algorithm. Time spent in sedentary, light and moderate-vigorous (MVPA) activity was established using published cut-points. Appropriate statistical techniques were utilised to account for the closed nature of the data (24-hour periods).

Results

Time spent asleep was related more to sedentary or light activity and not to MVPA. The most active (95^th percentile) children spent 55–84 fewer minutes asleep and 16–19 more minutes awake at night compared to the least active (5^th percentile) children. Children with later bedtimes slept less at night (30–40 minutes) and undertook more sedentary (10–15 minutes) but also more light (18–23 minutes) activity during the day. However, no differences in MVPA were apparent according to bedtime. Children slept slightly less on weekend nights (11 minutes) compared with week-nights, but only at 3 years of age. Most relationships were broadly similar at 3, 5 and 7 years of age.

Conclusion

Children who are more physically active during the day have shorter total sleep time and are more awake at night than less active children. The protective effect of sleep on obesity does not appear to be mediated by increased physical activity.     

How Do Thermophilic Proteins and Proteomes Withstand High Temperature?

We attempt to understand the origin of enhanced stability in thermophilic proteins by analyzing thermodynamic data for 116 proteins, the largest data set achieved to date. We compute changes in entropy and enthalpy at the convergence temperature where different driving forces are maximally decoupled, in contrast to the majority of previous studies that were performed at the melting temperature. We find, on average, that the gain in enthalpy upon folding is lower in thermophiles than in mesophiles, whereas the loss in entropy upon folding is higher in mesophiles than in thermophiles. This implies that entropic stabilization may be responsible for the high melting temperature, and hints at residual structure or compactness of the denatured state in thermophiles. We find a similar trend by analyzing a homologous set of proteins classified based only on the optimum growth temperature of the organisms from which they were extracted. We find that the folding free energy at the temperature of maximal stability is significantly more favorable in thermophiles than in mesophiles, whereas the maximal stability temperature itself is similar between these two classes. Furthermore, we extend the thermodynamic analysis to model the entire proteome. The results explain the high optimal growth temperature in thermophilic organisms and are in excellent quantitative agreement with full thermal growth rate data obtained in a dozen thermophilic and mesophilic organisms.     

Do Babies Resemble Their Fathers More Than Their Mothers? A Failure to Replicate Christenfeld and Hill (1995)

Contrary to Christenfeld and Hill (1995), we find that children aged 1, 3, and 5 do not appear to resemble their fathers significantly more than their mothers. We provide an explanation as to why this should be. In addition, we note that any father–child resemblance that does exist, although better than chance, is far from overwhelming.     

Do Women Prefer More Complex Music around Ovulation?

The evolutionary origins of music are much debated. One theory holds that the ability to produce complex musical sounds might reflect qualities that are relevant in mate choice contexts and hence, that music is functionally analogous to the sexually-selected acoustic displays of some animals. If so, women may be expected to show heightened preferences for more complex music when they are most fertile. Here, we used computer-generated musical pieces and ovulation predictor kits to test this hypothesis. Our results indicate that women prefer more complex music in general; however, we found no evidence that their preference for more complex music increased around ovulation. Consequently, our findings are not consistent with the hypothesis that a heightened preference/bias in women for more complex music around ovulation could have played a role in the evolution of music. We go on to suggest future studies that could further investigate whether sexual selection played a role in the evolution of this universal aspect of human culture.