Wolbachia Symbiont Infections Induce Strong Cytoplasmic Incompatibility in the Tsetse Fly Glossina morsitans

Tsetse flies are vectors of the protozoan parasite African trypanosomes, which cause sleeping sickness disease in humans and nagana in livestock. Although there are no effective vaccines and efficacious drugs against this parasite, vector reduction methods have been successful in curbing the disease, especially for nagana. Potential vector control methods that do not involve use of chemicals is a genetic modification approach where flies engineered to be parasite resistant are allowed to replace their susceptible natural counterparts, and Sterile Insect technique (SIT) where males sterilized by chemical means are released to suppress female fecundity. The success of genetic modification approaches requires identification of strong drive systems to spread the desirable traits and the efficacy of SIT can be enhanced by identification of natural mating incompatibility. One such drive mechanism results from the cytoplasmic incompatibility (CI) phenomenon induced by the symbiont Wolbachia. CI can also be used to induce natural mating incompatibility between release males and natural populations. Although Wolbachia infections have been reported in tsetse, it has been a challenge to understand their functional biology as attempts to cure tsetse of Wolbachia infections by antibiotic treatment damages the obligate mutualistic symbiont (Wigglesworthia), without which the flies are sterile. Here, we developed aposymbiotic (symbiont-free) and fertile tsetse lines by dietary provisioning of tetracycline supplemented blood meals with yeast extract, which rescues Wigglesworthia-induced sterility. Our results reveal that Wolbachia infections confer strong CI during embryogenesis in Wolbachia-free (GmmApo) females when mated with Wolbachia-infected (GmmWt) males. These results are the first demonstration of the biological significance of Wolbachia infections in tsetse. Furthermore, when incorporated into a mathematical model, our results confirm that Wolbachia can be used successfully as a gene driver. This lays the foundation for new disease control methods including a population replacement approach with parasite resistant flies. Alternatively, the availability of males that are reproductively incompatible with natural populations can enhance the efficacy of the ongoing sterile insect technique (SIT) applications by eliminating the need for chemical irradiation

Four Essays on Economic Evolution : an introduction

This essay is in two parts. The first considers the evolution of evolutionary economics from 1982 to 2012. While enormous advances are acknowledged, it is argued that the field is in danger of fragmentation and that there has been relatively little development in its over-arching theoretical framework since Nelson and Winter (1982). This sets the scene for a 2011 workshop and four of the papers presented at the event. In the second part, each paper is outlined in turn, both in terms of its specific contribution and any light it may shine on the problems raised in the first part.Peer reviewe

Technical Normativity

Despite their increasingly expanding empire, technical norms have scarcely attracted the attention of legal scholars. This article tries to understand the working of such norms by approaching the issue from the standpoint of the notion of \u201cartefactual normativity\u201d\u2014i.e. normativity concerning artefacts and their charac- teristics\u2014and then by analysing a strikingly growing class of technical norms more or less explicitly directed at influencing behaviour. What emerges from this analysis is a non-legal form of normativity which involves the subject in a process of con- tinuous observation, surveillance and direction of his/her conduct whose final aim is his/her self-identification with certain imposed standards. The paper ends with a section on the standardisation of the language of regulation

Defining the TRiC/CCT interactome links chaperonin function to stabilization of newly made proteins with complex topologies

Folding within the crowded cellular milieu often requires assistance from molecular chaperones that prevent inappropriate interactions leading to aggregation and toxicity. The contribution of individual chaperones to folding the proteome remains elusive. We here demonstrate that the eukaryotic chaperonin TRiC/CCT (TCP1-Ring Complex or Chaperonin Containing TCP1) has broad binding specificity in vitro similar to the prokaryotic chaperonin GroEL. However, in vivo TRiC substrate selection is not based solely on intrinsic determinants; instead, specificity is dictated by factors present during protein biogenesis. The identification of cellular substrates revealed that TRiC interacts with folding intermediates of a subset of structurally and functionally diverse polypeptides. Bioinformatics analysis revealed an enrichment in multidomain proteins and regions of beta strand propensity that are predicted to be slow-folding and aggregation-prone. Thus, TRiC may have evolved to protect complex protein topologies within its central cavity during biosynthesis and folding

Human TRiC complex purified from HeLa cells contains all eight CCT subunits and is active in vitro

Archaeal and eukaryotic cytosols contain group II chaperonins, which have a double-barrel structure and fold proteins inside a cavity in an ATP-dependent manner. The most complex of the chaperonins, the eukaryotic TCP-1 ring complex (TRiC), has eight different subunits, chaperone containing TCP-1 (CCT1–8), that are arranged so that there is one of each subunit per ring. Aspects of the structure and function of the bovine and yeast TRiC have been characterized, but studies of human TRiC have been limited. We have isolated and purified endogenous human TRiC from HeLa suspension cells. This purified human TRiC contained all eight CCT subunits organized into double-barrel rings, consistent with what has been found for bovine and yeast TRiC. The purified human TRiC is active as demonstrated by the luciferase refolding assay. As a more stringent test, the ability of human TRiC to suppress the aggregation of human γD-crystallin was examined. In addition to suppressing off-pathway aggregation, TRiC was able to assist the refolding of the crystallin molecules, an activity not found with the lens chaperone, α-crystallin. Additionally, we show that human TRiC from HeLa cell lysate is associated with the heat shock protein 70 and heat shock protein 90 chaperones. Purification of human endogenous TRiC from HeLa cells will enable further characterization of this key chaperonin, required for the reproduction of all human cells.National Institutes of Health (U.S.) (NIH Roadmap grant EY016525)National Eye Institute (NEI grant EY015834