Phosphatase localization in bacterial chemotaxis: divergent mechanisms, convergent principles

Chemotaxis is the process by which cells sense changes in their chemical environment and move towards more favorable conditions. In divergent species of bacteria, the chemotaxis proteins localize to the poles of the cell and information is transferred to the flagellar motors through the phosphorylation of a soluble protein CheY. Using mathematical models and computer simulation, we demonstrate that phosphatase localization controls the spatial distribution of CheY-P in the cytosol at steady state. Remarkably, the location of the phosphatase is not conserved in different species of bacteria. The sole phosphatase in Escherichia coli is localized with the signaling complex and the primary phosphatase in Bacillus subtilis is localized at the flagellar motors. Despite these alternate pathway structures, both designs minimize differences in the concentration of phosphorylated CheY proximal to each motor unlike a design where the phosphatase is freely diffusing in the cytoplasm. These results suggest that motile bacteria have evolved alternate mechanisms to ensure that each motor receives roughly the same signal at steady state. The hypothesis is that complex networks have evolved to satisfy certain design principles in order to function robustly. While specific mechanisms are different, the underlying principles of phosphatase localization in E. coli and B. subtilis appear to be the same.     

Left-right patterning: conserved and divergent mechanisms

The left-right (LR) asymmetry of visceral organs is fundamental to their function and position within the body. Over the past decade or so, the molecular mechanisms underlying the establishment of such LR asymmetry have been revealed in many vertebrate and invertebrate model organisms. These studies have identified a gene network that contributes to this process and is highly conserved from sea urchin to mouse. By contrast, some specific steps of the process, such as the symmetry-breaking event and situs-specific organogenesis, appear to have diverged during evolution. Here, we summarize the common and divergent mechanisms by which LR asymmetry is established in vertebrates.     

Normal brain ageing: models and mechanisms

Normal ageing is associated with a degree of decline in a number of cognitive functions. Apart from the issues raised by the current attempts to expand the lifespan, understanding the mechanisms and the detailed metabolic interactions involved in the process of normal neuronal ageing continues to be a challenge. One model, supported by a significant amount of experimental evidence, views the cellular ageing as a metabolic state characterized by an altered function of the metabolic triad: mitochondria-reactive oxygen species (ROS)-intracellular Ca2+. The perturbation in the relationship between the members of this metabolic triad generate a state of decreased homeostatic reserve, in which the aged neurons could maintain adequate function during normal activity, as demonstrated by the fact that normal ageing is not associated with widespread neuronal loss, but become increasingly vulnerable to the effects of excessive metabolic loads, usually associated with trauma, ischaemia or neurodegenerative processes. This review will concentrate on some of the evidence showing altered mitochondrial function with ageing and also discuss some of the functional consequences that would result from such events, such as alterations in mitochondrial Ca2+ homeostasis, ATP production and generation of ROS.     

The entomopathogenic bacterial endosymbionts Xenorhabdus and Photorhabdus: convergent lifestyles from divergent genomes

Members of the genus Xenorhabdus are entomopathogenic bacteria that associate with nematodes. The nematode-bacteria pair infects and kills insects, with both partners contributing to insect pathogenesis and the bacteria providing nutrition to the nematode from available insect-derived nutrients. The nematode provides the bacteria with protection from predators, access to nutrients, and a mechanism of dispersal. Members of the bacterial genus Photorhabdus also associate with nematodes to kill insects, and both genera of bacteria provide similar services to their different nematode hosts through unique physiological and metabolic mechanisms. We posited that these differences would be reflected in their respective genomes. To test this, we sequenced to completion the genomes of Xenorhabdus nematophila ATCC 19061 and Xenorhabdus bovienii SS-2004. As expected, both Xenorhabdus genomes encode many anti-insecticidal compounds, commensurate with their entomopathogenic lifestyle. Despite the similarities in lifestyle between Xenorhabdus and Photorhabdus bacteria, a comparative analysis of the Xenorhabdus, Photorhabdus luminescens, and P. asymbiotica genomes suggests genomic divergence. These findings indicate that evolutionary changes shaped by symbiotic interactions can follow different routes to achieve similar end points.     

Rho GTPases regulate axon growth through convergent and divergent signaling pathways

Rho GTPases are essential regulators of cytoskeletal reorganization, but how they do so during neuronal morphogenesis in vivo is poorly understood. Here we show that the actin depolymerization factor cofilin is essential for axon growth in Drosophila neurons. Cofilin function in axon growth is inhibited by LIM kinase and activated by Slingshot phosphatase. Dephosphorylating cofilin appears to be the major function of Slingshot in regulating axon growth in vivo. Genetic data provide evidence that Rho or Rac/Cdc42, via effector kinases Rok or Pak, respectively, activate LIM kinase to inhibit axon growth. Importantly, Rac also activates a Pak-independent pathway that promotes axon growth, and different RacGEFs regulate these distinct pathways. These genetic analyses reveal convergent and divergent pathways from Rho GTPases to the cytoskeleton during axon growth in vivo and suggest that different developmental outcomes could be achieved by biases in pathway selection.     

Neuropeptide binding reflects convergent and divergent evolution in species-typical group sizes

Neuroendocrine factors that produce species differences in aggregation behavior ("sociality") are largely unknown, although relevant studies should yield important insights into mechanisms of affiliation and social evolution. We here focused on five species in the avian family Estrildidae that differ selectively in their species-typical group sizes (all species are monogamous and occupy similar habitats). These include two highly gregarious species that independently evolved coloniality; two territorial species that independently evolved territoriality; and an intermediate, modestly gregarious species that is a sympatric congener of one of the territorial species. Using males and females of each species, we examined binding sites for (125)I-vasoactive intestinal polypeptide (VIP), (125)I-sauvagine (SG; a ligand for corticotropin releasing factor, CRF, receptors) and a linear (125)I-V(1a) vasopressin antagonist (to localize receptors for vasotocin, VT). VIP, CRF and VT are neuropeptides that influence stress, anxiety and/or various social behaviors. For numerous areas (particularly within the septal complex), binding densities in the territorial species differed significantly from binding in the more gregarious species, and in most of these cases, binding densities for the moderately gregarious species were either comparable to the two colonial species or were intermediate to the territorial and colonial species. Such patterns were observed for (125)I-VIP binding in the medial bed nucleus of the stria terminalis, medial septum, septohippocampal septum, and subpallial zones of the lateral septum; for (125)I-SG binding in the infundibular hypothalamus, and lateral and medial divisions of the ventromedial hypothalamus; and for the linear (125)I-V(1a) antagonist in the medial septum, and the pallial and subpallial zones of the caudal lateral septum. With the exception of (125)I-SG binding in the infundibular hypothalamus, binding densitites are positively related to sociality.     

Internal consistency and convergent and divergent validity of the Liverpool jetlag questionnaire

ABSTRACT

Objective measures of circadian disruption are difficult to capture in a free-living environment hence the importance of validating subjective measures of jetlag. We aimed to assess the internal consistency of the 15-item Liverpool Jetlag Scale and its convergent and divergent validity with indicators of fatigue and anxiety in a large sample of air passengers. Online survey of passengers was conducted after travel on a range of long-haul flights. Jetlag was captured using the Liverpool scale, fatigue was measured using the Vitality subscale of the Short-Form Health Survey (SF-36), and the presence of anxiety or worry before, during, and after flight was self-reported. Inter-item correlations and Cronbach’s alpha were calculated to assess the internal consistency of the scale. Exploratory factor analysis was used to examine whether the scale was consistent with one underlying construct of circadian disruption. Correlations between fatigue and anxiety (flying, situational, symptoms) with jetlag were used to assess convergent and divergent validity. Linear regression was used to determine the most important symptoms contributing to subjective jetlag rating. N = 460 passengers (57% female, mean age 50, SD 16 years) were surveyed. Cronbach’s alpha indicated high internal reliability (alpha = 0.85). Jetlag was more strongly correlated with fatigue (rho = 0.47) than any type of anxiety (rho = 0.10–0.22). Exploratory factor analysis indicated responses were consistent with four factors: (i) fatigue/daytime impairment, (ii) sleep disturbance, (iii) changes in appetite and (iv) changes in bowel function. Regression analysis indicated that only changes in concentration, sleep time, fatigue, sleep quality and frequency of bowel motions were independent correlates of subjective jetlag (R² = 27%). The Liverpool Jetlag Scale is internally consistent and demonstrates the expected relationships with fatigue and anxiety. Patterns of response are not consistent with all items being derived from one underlying factor, i.e. circadian disruption. Further, not all items contributed to the jetlag rating, suggesting the single-item rating may be useful for capturing the subjective experience of jetlag, whilst a total jetlag score is useful for also capturing circadian symptoms considered by passengers to be unrelated to jetlag. Validation of subjective jetlag against objective measures of circadian disruption is required.     

Cellular ageing mechanisms in osteoarthritis

Age is the strongest independent risk factor for the development of osteoarthritis (OA) and for many years this was assumed to be due to repetitive microtrauma of the joint surface over time, the so-called ‘wear and tear’ arthritis. As our understanding of OA pathogenesis has become more refined, it has changed our appreciation of the role of ageing on disease. Cartilage breakdown in disease is not a passive process but one involving induction and activation of specific matrix-degrading enzymes; chondrocytes are exquisitely sensitive to changes in the mechanical, inflammatory and metabolic environment of the joint; cartilage is continuously adapting to these changes by altering its matrix. Ageing influences all of these processes. In this review, we will discuss how ageing affects tissue structure, joint use and the cellular metabolism. We describe what is known about pathways implicated in ageing in other model systems and discuss the potential value of targeting these pathways in OA.     

Co-repressor induced order and biotin repressor dimerization: a case for divergent followed by convergent evolution

BirA catalyzes the adenylation and subsequent covalent attachment of biotin to the biotin carboxyl carrier protein (BCCP). In the absence of apo-BCCP, biotin-5'-AMP acts as a co-repressor that induces BirA dimerization and binding to the bio operator to repress biotin biosynthesis. The crystal structures of apo-BirA, and BirA in complex with biotin have been reported. We here describe the 2.8A resolution crystal structure of BirA in complex with the co-repressor analog biotinol-5'-AMP. It was previously shown that the structure of apo-BirA is monomeric and that binding of biotin weakly induces a dimeric structure in which three disordered surface loops become organized to form the dimer interface. The structure of the co-repressor complex is also a dimer, clearly related to the BirA.biotin structure, but with several significant conformational changes. A hitherto disordered "adenylate binding loop" forms a well-defined structure covering the co-repressor. The co-repressor buttresses the dimer interface, resulting in improved packing and a 12 degrees change in the hinge-bending angle along the dimer interface relative to the BirA.biotin structure. This helps explain why the binding of the co-repressor is necessary to optimize the binding of BirA to the bioO operator. The structure reveals an unexpected use of the nucleotide-binding motif GXGXXG in binding adenylate and controlling the repressor function. Finally, based on structural analysis we propose that the class of adenylating enzymes represented by BirA, lipoate protein ligase and class II tRNA synthetases diverged early and were selected based on their ability to sequester co-factors or amino acid residues, and adenylation activity arose independently through functional convergence.     

The EEG spatial pattern and psychophysiological characteristics of divergent and convergent thinking in humans

The psychophysiological parameters of the spatial synchronization of bioelectric potentials during the nonverbal divergent, nonverbal convergent, verbal divergent and verbal convergent thinking have been analyzed. The relationships between the performances of tests for these thinking styles were different. The strongest significant correlation has been found between the productivities of the verbal divergent and verbal convergent thinking styles; the weakest correlation, between those of the nonverbal divergent and nonverbal convergent thinking styles. The thinking styles differ from one another in the topography of spatial synchronization of bioelectric potentials. They are the most pronounced during the two nonverbal tasks and the least pronounced during the two verbal tasks. The thinking styles also differ from one another in the degree of the enhancement of the coherence between bioelectric potentials during tasks compared to the baseline (with the eyes open). Some human psychophysiological characteristics facilitating the performance of divergent (creative) tasks have a negative effect on the performance of convergent (noncreative) tasks. The data are discussed in terms of the differences in the level of general activation and involvement of different types of information processing (simultaneous and successive).     

Comparative analysis of spatial EEG organization on models of non-verbal divergent and convergent thinking

Features of neurophysiological organization of two main thinking types playing different roles in creative processes, i.e., divergent and convergent were studied with participation of 30 right-handed male subjects at the age from 30 to 50 years. Two tests were presented: (1) creation of many visual images on the basis of two simple geometrical figures (the model of divergent thinking) and (2) classification of a figure element with one of the offered standard samples (convergent thinking). The number of created images or correctly classified elements for five minutes served a criterion of performance productivity. It was found that performance of the divergent test with high productivity (as compared to low productivity) was characterized by a greater increase in non-linear interactions between the cortical potentials, especially in the axis right frontal--left occipital areas. At the same time, under conditions of high productivity, the number of active narrow-frequency spectral-coherent EEG bands increased. The data confirm the notion of neurophysiological organization of creative processes, according to which creative processes require the intensification of retrieval operations (both conscious and unconscious), based on extensive interhemispheric interaction and involvement of a system of EEG coherent structures oscillating with different frequencies.     

Non-genomic convergent and divergent signalling of rapid responses to aldosterone and estradiol in mammalian colon

Studies from our laboratory have demonstrated rapid ( < 1 min) non-genomic activation of Na(+)-H(+) exchange, K(+) recycling, PKC activity and a PKC-dependent Ca(2+) entry through L-type Ca(2+) channels specifically by mineralocorticoids in distal colon. Aldosterone directly stimulates the activity of the PKC alpha isoform (but not PKC delta, PKC epsilon and PKC zeta) in a cell-free assay system containing only purified commercially available enzyme, appropriate substrate peptide, co-factors and lipid vesicles. The primary ion transport target of the non-genomic signal transduction cascade elicited by aldosterone in epithelia is the Na(+)-H(+) exchanger. In isolated colonic crypts, aldosterone produced a PKC alpha sensitive intracellular alkalinisation within 1 min of hormone addition. Intracellular alkalinisation upregulates an ATP-dependent K(+) channel, which is involved in K(+) recycling to maintain the electrical driving force for Na(+) absorption, while inhibiting a Ca(2+) -dependent K(+) channel, which generates the charge balance for Cl(-) secretion. The non-genomic response to aldosterone in distal colon appears to enhance the capacity for absorption while down-regulating the potential for secretion. We have also demonstrated rapid (< 1 min) non-genomic activation of Na(+)-H(+) exchange, K(+) recycling, PKC alpha activity, and a PKC delta- and PKA-dependent Ca(2+) entry through di-hydropyridine-blockable Ca(2+) channels specifically by 17beta-estradiol in distal colon. These rapid effects are female gender specific and are insensitive to inhibitors of the classical estrogen receptor (ER). 17 beta-Estradiol directly stimulated the activity of both PKC delta and PKC alpha (but not PKC epsilon or PKC zeta) in a cell-free assay system. E2 rapidly inhibited basolateral K(Ca) channel activity which would be expected to result in an acute inhibition of Cl(-) secretion. Physiological concentrations of E2 (0.1-10 nM) reduced both basal and secretagogue-induced Cl(-) secretion. This anti-secretory effect of E2 is sensitive to PKC inhibition, intracellular Ca(2+) chelation, and is female gender specific and insensitive to inhibitors of the classical ER. These observations link rapid non-genomic activation of second messengers with a rapid gender-specific physiological effect in the whole tissue. Aldosterone and E2 differ in their protein kinase signal transduction and both hormones stimulate specific PKC isoforms indicating both common and divergent signalling systems for salt-retaining steroid hormones. The physiological function of non-genomic effects of aldosterone and estradiol is to shift the balance from net secretion to net absorption in a pluripotential epithelium.     

Acute oxygen sensing: diverse but convergent mechanisms in airway and arterial chemoreceptors

Airway neuroepithelial bodies sense changes in inspired O₂, whereas arterial O₂ levels are monitored primarily by the carotid body. Both respond to hypoxia by initiating corrective cardiorespiratory reflexes, thereby optimising gas exchange in the face of a potentially deleterious O₂ supply. One unifying theme underpinning chemotransduction in these tissues is K⁺ channel inhibition. However, the transduction components, from O₂ sensor to K⁺ channel, display considerable tissue specificity yet result in analogous end points. Here we highlight how emerging data are contributing to a more complete understanding of O₂ chemosensing at the molecular level.     

How and why the mechanisms of ageing and carcinogenesis originated: a hypothesis

A hypothesis explaning the nature of ageing and carcinogenesis is proposed. The hypothesis is based on the notion of a unitary organism as a module of a colony that has lost the ability to reproduce asexually. The evolutionary forerunner of the old age involution was the asexual iteroparity of colony modules, and the evolutionary forerunner of carcinogenesis was the semelparity of such colonies; the evolutionary forerunner or tumour regression was the switch of the colony module from semelparity to iteroparity. The donor-acceptor relations of a colony module emerge in unitary organisms in the course of ageing and carcinogenesis. The mechanisms of ageing and carcinogenesis originated at the stages of Metazoa evolution when the organisms had not yet acquired the nervous and the endocrine systems.     

Cancer cachexia: the molecular mechanisms

Cancer cachexia is a syndrome characterised by a marked weight loss, anorexia, asthenia and anaemia. In fact, many patients who die with advanced cancer suffer from cancer cachexia. The cachectic state is invariably associated with the presence and growth of the tumour and leads to a malnutrition status due to the induction of anorexia or decreased food intake. In addition, the competition for nutrients between the tumour and the host leads to an accelerated starvation state which promotes severe metabolic disturbances in the host, including hypermetabolism which leads to an increased energetic inefficiency. Although, the search for the cachectic factor(s) started a long time ago, and although many scientific and economic efforts have been devoted to its discovery, we are still a long way from knowing the whole truth. The main aim of the present review is to summarise and evaluate the different catabolic mediators (both humoural and tumoural) involved in cancer cachexia since they may represent targets for future promising clinical investigations.