TCTP in Development and Cancer

The translationally controlled tumor protein (TCTP) is highly conserved among animal species. It is widely expressed in many different tissues. It is involved in regulating many fundamental processes, such as cell proliferation and growth, apoptosis, pluripotency, and the cell cycle. Hence, it is not surprising that it is essential for normal development and, if misregulated, can lead to cancer. Provided herein is an overview of the diverse functions of TCTP, with a focus on development. Furthermore, we discuss possible ways by which TCTP misregulation or mutation could result in cancer

Histone H3 lysine 4 methylation is associated with the transcriptional reprogramming efficiency of somatic nuclei by oocytes.

BACKGROUND: When the nuclei of mammalian somatic cells are transplanted to amphibian oocytes in the first meiotic prophase, they are rapidly induced to begin transcribing several pluripotency genes, including Sox2 and Oct4. The more differentiated the donor cells of the nuclei, the longer it takes for the pluripotency genes to be activated after the nuclear transfer to oocytes. We have used this effect in order to investigate the role of histone modifications in this example of nuclear reprogramming. RESULTS: Reverse transcription polymerase chain reaction analysis shows that the transcriptional reprogramming of pluripotency genes, such as Sox2 and Oct4, takes place in transplanted nuclei from C3H10T1/2 cells and from newly differentiated mouse embryonic stem cells. We find that the reprogramming of 10T1/2 nuclei is accompanied by an increased phosphorylation, an increased methylation and a rapidly reduced acetylation of several amino acids in H3 and other histones. These results are obtained by the immunofluorescent staining of transplanted nuclei and by Western blot analysis. We have also used chromatin immunoprecipitation analysis to define histone modifications associated with the regulatory or coding regions of pluripotency genes in transplanted nuclei. Histone phosphorylation is increased and histone acetylation is decreased in several regulatory and gene coding regions. An increase of histone H3 lysine 4 dimethylation (H3K4 me2) is seen in the regulatory regions and gene coding region of pluripotency genes in reprogrammed nuclei. Furthermore, histone H3 lysine 4 trimethylation (H3K4 me3) is observed more strongly in the regulatory regions of pluripotency genes in transplanted nuclei that are rapidly reprogrammed than in nuclei that are reprogrammed slowly and are not seen in beta-globin, a gene that is not reprogrammed. When 10T1/2 nuclei are incubated in Xenopus oocyte extracts, histone H3 serine 10 (H3S10) is strongly phosphorylated within a few hours. Immunodepletion of Aurora B prevents this phosphorylation. CONCLUSION: We conclude that H3K4 me2 and me3 are likely to be important for the efficient reprogramming of pluripotency genes in somatic nuclei by amphibian oocytes and that Aurora B kinase is required for H3S10 phosphorylation which is induced in transplanted somatic cell nuclei.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

The evolving biology of cell reprogramming

Modern stem cell biology has achieved a transformation that was thought by many to be every bit as unattainable as the ancient alchemists' dream of transforming base metals into gold. Exciting opportunities arise from the process known as ‘cellular reprogramming’ in which cells can be reliably changed from one tissue type to another. This is enabling novel approaches to more deeply investigate the fundamental basis of cell identity. In addition, new opportunities have also been created to study (perhaps even to treat) human genetic and degenerative diseases. Specific cell types that are affected in inherited disease can now be generated from easily accessible cells from the patient and compared with equivalent cells from healthy donors. The differences in cellular phenotype between the two may then be identified, and assays developed to establish therapies that prevent the development or progression of disease symptoms. Cellular reprogramming also has the potential to create new cells to replace those whose death or dysfunction causes disease symptoms. For patients suffering from inherited cases of degenerative diseases like Parkinson's disease or amyotrophic lateral sclerosis (also known as motor neuron disease), the future realization of such cell-based therapies would truly be worth its weight in gold. However, before this enormous potential can become a reality, several significant biological and technical challenges must be overcome. Furthermore, to maintain the credibility of the scientific community with the general public, it is important that hope-inspiring advances are not over-hyped. The papers in this issue of the Philosophical Transactions of the Royal Society B: Biological Sciences cover many areas relevant to this topic. In this Introduction, we provide an overall context in which to consider these individual papers

An indelible lineage marker for Xenopus using a mutated green fluorescent protein

We describe the use of a DNA construct (named GFP.RN3) encoding green fluorescent protein as a lineage marker for Xenopus embryos. This offers the following advantages over other lineage markers so far used in Xenopus. When injected as synthetic mRNA, its protein emits intense fluorescence in living embryos. It is non-toxic, and the fluorescence does not bleach when viewed under 480 nm light. It is surprisingly stable, being strongly visible up to the feeding tadpole stage (5 days), and in some tissues for several weeks after mRNA injection. We also describe a construct that encodes a blue fluorescent protein. We exemplify the use of this GFP.RN3 construct for marking the lineage of individual blastomeres at the 32- to 64-cell stage, and as a marker for single transplanted blastula cells. Both procedures have revealed that the descendants of one embryonic cell can contribute single muscle cells to nearly all segmental myotomes rather than predominantly to any one myotome. An independent aim of our work has been to follow the fate of cells in which an early regulatory gene has been temporarily overexpressed. For this purpose, we co-injected GFP.RN3 mRNA and mRNA for the early Xenopus gene Eomes, and found that a high concentration of Eomes results in ectopic muscle gene activation in only the injected cells. This marker may therefore be of general value in providing long term identification of those cells in which an early gene with ephemeral expression has been overexpressed

TCTP in development and cancer.

The translationally controlled tumor protein (TCTP) is highly conserved among animal species. It is widely expressed in many different tissues. It is involved in regulating many fundamental processes, such as cell proliferation and growth, apoptosis, pluripotency, and the cell cycle. Hence, it is not surprising that it is essential for normal development and, if misregulated, can lead to cancer. Provided herein is an overview of the diverse functions of TCTP, with a focus on development. Furthermore, we discuss possible ways by which TCTP misregulation or mutation could result in cancer.Peer Reviewe

Collective Bargaining in the Maritime Fishing Industry: Recent Developments

La débâcle de l'industrie de la pêche en 1974 et l'extension de la compétence canadienne à la limite de 200 milles des cotes en janvier 1977 ont été les événements majeurs qui ont conduit à une participation plus active de la part du gouvernement fédéral. Des quotas et une politique de permis plus rigoureuse ont été instaures, le ministère des Pêcheries fut décentralise par la nomination d'administrateurs régionaux et de fonctionnaires préposes aux services communautaires afin de permettre aux pêcheurs d'obtenir un rendement accru au niveau local et, enfin, le ministre Leblanc encouragea la syndicalisation comme moyen de donner une voix plus forte aux pêcheurs dans les questions se rapportant à l'industrie. Le Syndicat des pêcheurs des Maritimes (Maritime Fishermen's Union) fut forme en mars 1977 dans le but précis de syndicaliser les pêcheurs côtiers. Le Syndicat obtint l'adhésion de 900 des 1 200 capitaines à plein temps entre Caraquet et Cape Tormentine et il implanta des sections locales dans les trois provinces Maritimes. Il réussit aussi à signer quelques ententes de gré à gré en matière de fixation des prix, mais ses efforts portèrent surtout sur l'obtention d'une législation relative à la négociation collective. De fait, la Loi sur la négociation collective dans l'industrie de la pêche au Nouveau-Brunswick (New Brunswick Fisheries Bargaining Act) fut promulguée en septembre 1982. Elle s'inspirait de la Loi sur les relations du travail provinciale (Industrial Relations Act), quoiqu'une de ses caractéristiques inusitées soit l'établissement obligatoire d'unités de négociation par entité géographique correspondant aux zones d'administration du ministère des Pêcheries (Nord-ouest du Nouveau-Brunswick, Sud-est du Nouveau-Brunswick et la baie de Fundy), et aussi par espèce de poisson. Le Syndicat a demande l'accréditation pour représenter les pêcheurs de sept espèces différentes dans le Nord-est et de neuf espèces dans le Sud-est. Des stipulations juridiques relatives à la situation de dépendance des pêcheurs ont longtemps prive les associations de pêcheurs de négocier au nom de leurs membres parce que, considères comme des commerçants aptes à vendre leurs prises à n'importe quel acheteur, ils ont toujours été exposes à l'accusation de restreindre le commerce en vertu de la Loi relative aux enquêtes sur les coalitions (Combines Investigation Act). Pour déterminer s'il s'agissait là d'un fait juridique ou d'une simple fiction, on peut soumettre les constatations révélées par une enquête faite en 1902 chez tous les capitaines de bateaux côtiers inscrits au répertoire du gouvernement du Nouveau-Brunswick. Le questionnaire a été adresse tant en français qu'en anglais et 552 d'entre eux furent remplis et considères comme valables. Il en ressort ce qui suit. 83 pour cent d'entre eux exerçaient le métier de pêcheurs depuis au moins deux générations, 95 pour cent étaient engages dans la pratique de la pêche depuis cinq ans ou plus, 23 pour cent avaient travaille à un titre quelconque dans une usine de traitement de poisson et 46 pourcent etaient des pêcheurs maries et ont déclare que leur épouse travaillait dans cette industrie, 216 etaient membres du Syndicat, 126 d'une coopérative locale et 168 d'une association quelconque. Quelques-uns étaient membres de plusieurs groupements et 119 indiquaient qu'ils n'appartenaient à aucune organisation. La région du Nord-est a fourni 271 réponses, le Sud-est, 209 et la baie de Fundy, 72. Une série de questions traitaient du type de relations avec les acheteurs. 31 pour cent avaient l'impression que les pêcheurs étaient subordonnes aux entreprises au niveau local, 23 pour cent estimaient que c'était les entreprises qui dépendaient des pêcheurs, tandis que 46 pour cent considéraient que leurs rapports étaient équilibres. Dans la région de la baie de Fundy, on était plus d'accord sur l'opinion qu'il y avait égalité de pouvoir entre les deux groupes, en partie parce que la saison de pêche est plus longue et parce que un plus grand nombre d'espèces utilisables sont disponibles, en partie parce qu'on y ressent moins le poids de l'exploitation passée. De plus, tandis qu'il existait un grand mécontentement touchant la façon dont les prix sont fixes d'une manière générale, cette insatisfaction était moins marquée dans la région de la baie de Fundy. En réalité, les sentiments d'opposition envers le gouvernement et le syndicat semblaient être plus prononces que l'antagonisme contre les entreprises. Enfin, les pêcheurs qui n'étaient pas membres du syndicat donnaient l'impression d'une plus grande dépendance que ceux qui en faisaient partie. Toutefois, l'étude ne permet pas de conclure que l'appartenance à une organisation renforce le sentiment de contrôle. Le contraire peut être aussi vrai. Une deuxième série de questions portaient sur la perception que se font les pêcheurs de leur propre image. En résume, ils se considèrent comme des propriétaires qui accomplissent un travail manuel pénible. Une minorité importante d'entre eux estiment qu'ils font partie de la classe ouvrière, bien que plusieurs notent une amélioration récente de leur image dans la population et de leur statut professionnel (surtout chez les Acadiens) et considèrent le recours à la grève inapproprié et impossible. Une des conséquences en est que le Syndicat doit modifier sa rhétorique radicale et continuer à s'impliquer activement comme partenaire important dans la gestion de l'industrie. D'autre part, les dirigeants d'associations et les gérants de coopératives ont besoin d'être sensibilises au fait que beaucoup de leurs membres conçoivent le monde et l'industrie selon l'esprit de classe et que, en conséquence, l'utilisation des termes «professionnalisme» et de «petite entreprise» peut s'avérer illusoire. On peut conclure que cette enquête, la première du genre, fait assez ressortir un sentiment de dépendance parmi les pêcheurs et une prise de conscience suffisante de leurs intérêts distincts pour justifier le gouvernement de développer les droits de négociation. D'autres provinces côtières devaient étudier soigneusement l'expérience de la loi du Nouveau-Brunswick afin de mettre au point leur propre législation.This paper summarizes recent changes in the Canadian fishing industry, with particular emphasis on the Maritime region, traces the rise of a new union seeking to represent inshore fishermen, and describes the resulting collective bargaining legislation in New Brunswick

Dynamic Interpretation of Hedgehog Signaling in the Drosophila Wing Disc

Morphogens are classically defined as molecules that control patterning by acting at a distance to regulate gene expression in a concentration-dependent manner. In the Drosophila wing imaginal disc, secreted Hedgehog (Hh) forms an extracellular gradient that organizes patterning along the anterior–posterior axis and specifies at least three different domains of gene expression. Although the prevailing view is that Hh functions in the Drosophila wing disc as a classical morphogen, a direct correspondence between the borders of these patterns and Hh concentration thresholds has not been demonstrated. Here, we provide evidence that the interpretation of Hh signaling depends on the history of exposure to Hh and propose that a single concentration threshold is sufficient to support multiple outputs. Using mathematical modeling, we predict that at steady state, only two domains can be defined in response to Hh, suggesting that the boundaries of two or more gene expression patterns cannot be specified by a static Hh gradient. Computer simulations suggest that a spatial “overshoot” of the Hh gradient occurs, i.e., a transient state in which the Hh profile is expanded compared to the Hh steady-state gradient. Through a temporal examination of Hh target gene expression, we observe that the patterns initially expand anteriorly and then refine, providing in vivo evidence for the overshoot. The Hh gene network architecture suggests this overshoot results from the Hh-dependent up-regulation of the receptor, Patched (Ptc). In fact, when the network structure was altered such that the ptc gene is no longer up-regulated in response to Hh-signaling activation, we found that the patterns of gene expression, which have distinct borders in wild-type discs, now overlap. Our results support a model in which Hh gradient dynamics, resulting from Ptc up-regulation, play an instructional role in the establishment of patterns of gene expression

H3K4 Methylation-Dependent Memory of Somatic Cell Identity Inhibits Reprogramming and Development of Nuclear Transfer Embryos

Vertebrate eggs can induce the nuclear reprogramming of somatic cells to enable production of cloned animals. Nuclear reprogramming is relatively inefficient, and the development of the resultant embryos is frequently compromised, in part due to the inappropriate expression of genes previously active in the donor nucleus. Here, we identify H3K4 methylation as a major epigenetic roadblock that limits transcriptional reprogramming and efficient nuclear transfer (NT). Widespread expression of donor-cell-specific genes was observed in inappropriate cell types in NT embryos, limiting their developmental capacity. The expression of these genes in reprogrammed embryos arises from epigenetic memories of a previously active transcriptional state in donor cells that is characterized by high H3K4 methylation. Reducing H3K4 methylation had little effect on gene expression in donor cells, but it substantially improved transcriptional reprogramming and development of NT embryos. These results show that H3K4 methylation imposes a barrier to efficient nuclear reprogramming and suggest approaches for improving reprogramming strategies.This work was funded by the Molecular Research Council ( MR/P000479/1 ), the Wellcome Trust ( 101050/Z/13/Z and 092096/Z/10/Z ), and Cancer Research UK ( C6946/A14492 ). E.H. was a recipient of a long-term fellowship from the European Molecular Biology Organization (EMBO) and Isaac Newton Trust funding

Rising influence of synthetic biology in regenerative medicine

This is an Open Access Article. It is published by IET under the Creative Commons Attribution 3.0 Unported Licence (CC BY). Full details of this licence are available at: http://creativecommons.org/licenses/by/3.0/Synthetic biology is an emerging area of research that combines the investigative nature of biology with the constructive nature of engineering. Despite the field being in its infancy, it has already aided the development of a myriad of industrially and pharmaceutically useful compounds, devices and therapies and is now being applied within the field of regenerative medicine. By combining synthetic biology with regenerative medicine, the engineering of cells and organisms offers potential avenues for applications in tissue engineering, bioprocessing, biomaterial and scaffold development, stem cell therapies and even gene therapies. This review aims to discuss how synthetic biology has been applied within these distinct areas of regenerative medicine, the challenges it faces and any future possibilities this exciting new field may hold