The TMEM16 Protein Family: A New Class of Chloride Channels?

Cl⁻ channels play important roles in many physiological processes, including transepithelial ion absorption and secretion, smooth and skeletal muscle contraction, neuronal excitability, sensory perception, and cell volume regulation. The molecular identity of many types of Cl⁻ channels is still unknown. Recently, three research groups have arrived independently at the identification of TMEM16A (also known as anoctamin-1) as a membrane protein strongly related to the activity of Ca²⁺-activated Cl⁻ channels (CaCCs). Site-specific mutagenesis of TMEM16A alters the properties of the channels, thus suggesting that TMEM16A forms, at least in part, the CaCC. TMEM16A is a member of a family that includes nine other membrane proteins. All TMEM16 proteins have a similar structure, with eight putative transmembrane domains and cytosolic amino- and carboxy-termini. TMEM16B expression also evokes the appearance of CaCCs, but with biophysical characteristics (voltage dependence, unitary conductance) different from those associated to TMEM16A. The roles of the other TMEM16 proteins are still unknown. The study of TMEM16 proteins may lead to identification of novel molecular mechanisms underlying ion transport and channel gating by voltage and Ca²⁺.     

The Molecular Basis of Bacterial–Insect Symbiosis

Insects provide experimentally tractable and cost-effective model systems to investigate the molecular basis of animal–bacterial interactions. Recent research is revealing the central role of the insect innate immune system, especially anti-microbial peptides and reactive oxygen species, in regulating the abundance and composition of the microbiota in various insects, including Drosophila and the mosquitoes Aedes and Anopheles. Interactions between the immune system and microbiota are, however, bidirectional with evidence that members of the resident microbiota can promote immune function, conferring resistance to pathogens and parasites by both activation of immune effectors and production of toxins. Antagonistic and mutualistic interactions among bacteria have also been implicated as determinants of the microbiota composition, including exclusion of pathogens, but the molecular mechanisms are largely unknown. Some bacteria are crucial for insect nutrition, through provisioning of specific nutrients (e.g., B vitamins, essential amino acids) and modulation of the insect nutritional sensing and signaling pathways (e.g., insulin signaling) that regulate nutrient allocation, especially to lipid and other energy reserves. A key challenge for future research is to identify the molecular interaction between specific bacterial effectors and animal receptors, as well as to determine how these interactions translate into microbiota-dependent signaling, metabolism, and immune function in the host.     

The Dicistroviridae： An Emerging Family of Invertebrate Viruses

Dicistroviruses comprise a newly characterized and rapidly expanding family of small RNA viruses of invertebrates. Several features of this virus group have attracted considerable research interest in recent years. In this review I provide an overview of the Dicistroviridae and describe progress made toward the understanding and practical application of dicistroviruses, including (i) construction of the first infectious clone of a dicistrovirus, (ii) use of the baculovirus expression system for production of an infectious dicistrovirus, (iii) the use of Drosophila C virus for analysis of host response to virus infection, and (iv) correlation of the presence of Israeli acute paralysis virus with honey bee colony collapse disorder. The potential use of dicistroviruses for insect pest management is also discussed. The structure, mechanism and practical use of the internal ribosome entry site (IRES) elements has recently been reviewed elsewhere.     

The Role of Altered Cell–Cell Communication in Melanoma Progression

Under normal homeostasis, melanocyte growth and behaviour is tightly controlled by the surrounding keratinocytes. Keratinocytes regulate melanocyte behaviour through a complex system of paracrine growth factors and cell-cell adhesion molecules. Pathological changes, leading to development of malignant melanoma, upset this delicate homeostatic balance and can lead to altered expression of cell-cell adhesion and cell-cell communication molecules. In particular, there is a switch from the E-cadherin-mediated keratinocyte-melanocyte partnership to the N-cadherin-mediated melanoma-melanoma and melanoma-fibroblast interaction. Other changes include the alteration in the gap junctions formed between the melanocyte and keratinocyte. Changes in the connexin expression, in particular the loss of connexin 43, may result in a reduction or a loss of gap junctional activity, which is thought to contribute towards tumour progression. In the current review we describe the alterations in cell-cell adhesion and communication associated with melanoma development and progression, and discuss how a greater understanding of these processes may aid the future therapy of this disease.     

Leaders in Cell Adhesion: An Interview with Richard Hynes,Pioneer of Cell–Matrix Interactions

Abstract

On a recent visit Richard O Hynes, FRS, HHMI, Daniel K. Ludwig Professor for Cancer Research at the Koch Institute for Integrative Cancer Research, MIT, graciously agreed to be interviewed in person for the first in Cell Communication and Adhesion's series on “Leaders in Cell Adhesion”. In this interview we discussed three things: 1) the early role of family, mentors, and luck on his career path; 2) his major discoveries of fibronectin, integrins and the evolution of extracellular matrix proteins; and 3) his role in, and thoughts on, current science policy. This interview reveals his characteristic calmness and infectious optimism, his spontaneous and down to earth sense of humor, and his great ability to place scientific questions in perspective. The interview, carried out on April 30^th 2013 is reported here verbatim with only minor editing for clarity.     

The CAR group of Ig cell adhesion proteins–Regulators of gap junctions?

Members of the CAR group of Ig-like type I transmembrane proteins mediate homotypic cell adhesion, share a common overall extracellular domain structure and are closely related at the amino acid sequence level. CAR proteins are often found at tight junctions and interact with intracellular scaffolding proteins, suggesting that they might modulate tight junction assembly or function. However, impairment of tight junction integrity has not been reported in mouse knockout models or zebrafish mutants of CAR members. In contrast, in the same knockout models deficits in gap junction communication were detected in several organ systems, including the atrioventricular node of the heart, smooth muscle cells of the intestine and the ureter and in Sertoli cells of the testes. Possible interactions between BT-IgSF and connexin41.8 on the disturbed pattern of pigment stripes found in zebrafish mutants and between ESAM and connexin43 during hematopoiesis in the mouse are also discussed. On the basis of the combined data and phenotypic similarities between CAR member mutants and connexin mutants I hypothesize that they primarily play a role in the organization of gap junction communication. Also see the video abstract here: https://youtu.be/i0yq2KhuDAE .     

The Role of Gap Junctions in Endothelial–Stromal Cell Interactions

Human Physiology - Gap junctions are one of the most highly specialized intercellular communications providing not only electrical coupling but also metabolic cooperation between cells due to the...     

The Evolution of Virulence in RNA Viruses under a Competition–Colonization Trade-Off

RNA viruses exist in large intra-host populations which display great genotypic and phenotypic diversity. We analyze a model of viral competition between two viruses infecting a constantly replenished cell pool. We assume a trade-off between the ability of the virus to colonize new cells (cell killing rate or virulence) and its local competitiveness (replicative success within coinfected cells). We characterize the conditions that allow for viral spread by means of the basic reproductive number and show that a local coexistence equilibrium exists, which is asymptotically stable. At this equilibrium, the less virulent competitor has a reproductive advantage over the more virulent colonizer reflected by a larger equilibrium population size of the competitor. The equilibria at which one virus outcompetes the other one are unstable, i.e., a second virus is always able to permanently invade. We generalize the two-virus model to multiple viral strains, each displaying a different virulence. To account for the large phenotypic diversity in viral populations, we consider a continuous spectrum of virulences and present a continuum limit of this multiple viral strains model that describes the time evolution of an initial continuous distribution of virulence without mutations. We provide a proof of the existence of solutions of the model equations, analytically assess the properties of stationary solutions, and present numerical approximations of solutions for different initial distributions. Our simulations suggest that initial continuous distributions of virulence evolve toward a distribution that is extremely skewed in favor of competitors. At equilibrium, only the least virulent part of the population survives. The discrepancy of this finding in the continuum limit with the two-virus model is attributed to the skewed equilibrium subpopulation sizes and to the transition to a continuum. Consequently, in viral quasispecies with high virulence diversity, the model predicts collective virulence attenuation. This result may contribute to understanding virulence attenuation, which has been reported in several experimental studies.     

The Scent of Symmetry: A Human Sex Pheromone that Signals Fitness?

A previous study by the authors showed that the body scent of men who have greater body bilateral symmetry is rated as more attractive by normally ovulating (non-pill-using) women during the period of highest fertility based on day within the menstrual cycle. Women in low-fertility phases of the cycle and women using hormone-based contraceptives do not show this pattern. The current study replicated these findings with a larger sample and statistically controlled for men's hygiene and other factors that were not controlled in the first study. The current study also examined women's scent attractiveness to men and found no evidence that men prefer the scent of symmetric women. We propose that the scent of symmetry is an honest signal of phenotypic and genetic quality in the human male, and chemical candidates are discussed. In both sexes, facial attractiveness (as judged from photos) appears to predict body scent attractiveness to the opposite sex. Women's preference for the scent associated with men's facial attractiveness is greatest when their fertility is highest across the menstrual cycle. The results overall suggest that women have an evolved preference for sires with good genes.     

The Body and Face of Woman: One Ornament that Signals Quality?

Evidence has accumulated in recent years supporting the hypothesis that both facial and bodily physical attractiveness in humans are certifications of developmental and hormonal health. Such evidence indicates that physical attractiveness is an honest or Zahavian signal of phenotypic and genetic quality. The hypothesis that physical beauty connotes health was first proposed by Westermarck and was discussed later by Ellis and Symons. It has been suggested that facial attractiveness in women is a deceptive signal of youth, unrelated to phenotypic and genetic quality. This sensory-bias or super-stimulus hypothesis is not supported by this study of men’s ratings of the attractiveness of photographs of 92 nude women. Independent ratings of photographs of faces, fronts with faces covered, and backs of the same women are significantly, positively correlated. The correlation between the ratings of different photos implies that women’s faces and external bodies comprise a single ornament of honest mate value, apparently constructed during puberty by estrogen and also probably by developmental adaptations for symmetry. Thus, women’s physical attractiveness in face and body honestly signal hormonal and perhaps developmental health.     

The grassland of Orkney: An œcological analysis

Summary

World-wide, our coastal waters have been subject to an increased nutrient input since the latter part of the nineteenth century. This has led to the eutrophication or ‘nutrient pollution’ of many coastal sites, including Langstone Harbour and the Ythan Estuary here in the UK. Eutrophication at these and, indeed, at other nutrient enriched sites is evident by the appearance of large blooms of fast-growing opportunistic macroalgae. Blooms of macroscopic species of green algae (Chlorophyta: Ulvophyceae) are particularly common and the phenomenon is often referred to as the occurrence of green tides.

Green tides may have a dramatic environmental impact, causing much damage to the local ecosystem. Numerous strategies have hence been employed in order to combat the problem, but to date there has been limited success. For this reason, current research in the UK is aimed at increasing our knowledge of green tide algae in terms of their ecophysiology, whilst further investigation of the nutrient pathways and fluxes within specific ecosystems has been deemed necessary. It is anticipated that this ‘backto basics’ approach will ultimately contribute to the development of new, successful eutrophication management strategies.     

The Study of Erwinia carotovora Bacteriocins with the Aid of Nalidixic Acid–Resistant Bacterial Indicator Cells

A novel approach is proposed for the study of the macromolecular bacteriocins of Erwinia carotovora (MCTVs). The approach lies in the bacteriocinogeny of pectolytic erwinia being studied using a lawn of a bacterial mutant resistant to nalidixic acid, an inducer of MCTVs. The high efficiency of this approach was demonstrated by studying carotovoricins in 104 different E. carotovora strains, 88% of which bear MCTVs, distinguished by the morphology of zones of induced lysis on a lawn of susceptible cells, the lysis pattern, and some other characteristics. Preliminary studies using this approach showed that there is no correlation between the occurrence of MCTVs in particular E. carotovora strains and the habitat of the host plants from which these strains were isolated. There are grounds to believe that the approach proposed can also be used for investigating bacterial lysogeny.     

An Evolution Model for Sequence Length Based on Residue Insertion–Deletion Independent of Substitution: An Application to the GC Content in Bacterial Genomes

We introduce here a gene evolution model which is an extension of the time-continuous stochastic IDIS model (Lèbre and Michel in J. Comput. Biol. Chem. 34:259-267, 2010) to sequence length. This new IDISL (Insertion Deletion Independent of Substitution based on sequence Length) model gives an analytical expression of the residue occurrence probability p(l) at sequence length l depending on stochastically independent processes of substitution, insertion, and deletion. Furthermore, in contrast to all mathematical models in this research field, the substitution, insertion, and deletion parameters of the IDISL model are independent of each other. For any diagonalizable substitution matrix M, the residue occurrence probability p(l) is given as a function of the eigenvalues of M, the eigenvector matrix of M, a vector r of the residue insertion rates, a deletion rate d (unlike our previous IDIS model), and a vector of the initial residue occurrence probability p(l(0)) at sequence length l(0).As another difference with the classical evolution approaches which mainly focus on sequence alignment, the IDIS class of models allows a mathematical analysis of the behavior of the residue occurrence probability according to either evolution time or sequence length. The length parameter can be associated with any nucleotide regions: genes, genomes, introns, repeats, 5' and 3' regions, etc. Three properties of the IDISL model are given in relation with the sequence length l: parameter scale, inverse evolution, and residue equilibrium distribution. Nucleotide occurrence probabilities are given in the particular case of the IDISL-HKY model, i.e. the IDISL model associated with the HKY asymmetric substitution matrix (Hasegawa et al. in J. Mol. Evol. 22:160-174, 1985).An application of the IDISL model is developed for a massive statistical analysis of GC content in all complete bacterial genomes available to date (894 non-anaerobic and anaerobic genomes). The IDISL-HKY model confirms the increase of the GC content with the genome length for two non-anaerobic taxonomic groups of bacterial genomes. Moreover, the non-linear modelling proposed by the IDISL model outperforms the most recent modelling of GC content in these bacterial genomes (Wang et al. in Biochem. Biophys. Res. Commun. 342:681-684, 2006; Musto et al. in Biochem. Biophys. Res. Commun. 347:1-3, 2006).     

The Production of a Stably Transformed Insect Cell Line Expressing An Invertebrate GABAA Receptor β-Subunit

Abstract

We have produced a stable insect cell line derived from Spodoptera frugiperda (Sf9) cells expressing a cDNA encoding a β-subunit of the Lymnaea stagnalis GABA_A receptor. The cDNA was randomly integrated into the insect cell genome under the control of a baculovirus immediate early gene (IE-1) promoter. Stable cell lines were established by transformation of Sf9 cells with the expression vector pIEK1.LGβ1 together with a plasmid encoding a selectable marker which confers neomycin (G418) resistance. Following growth in the presence of G418, neomycin resistant clones were selected, amplified and analysed for the presence of functional GABA-gated chloride channels. Electrophysiological analysis of one cell line showed the presence of a picrotoxin-sensitive chloride channel not present in control Sf9 cells. These channels were also sensitive to GABA, albeit at relatively high (mM) concentrations.