A versatile plasmid system for reconstitution and analysis of mammalian ubiquitination cascades in yeast

Ubiquitination is a posttranslational protein modification that regulates most aspects of cellular life. The sheer number of ubiquitination enzymes that are present in a mammalian cell, over 700 in total, has thus far hampered the analysis of distinct protein ubiquitination cascades in a cellular context. To overcome this complexity we have developed a versatile vector system that allows the reconstitution of specific ubiquitination cascades in the model eukaryote Saccharomyces cerevisae (baker’s yeast). The vector system consists of 32 modular yeast shuttle plasmids allowing inducible or constitutive expression of up to four proteins of interest in a single cell. To demonstrate the validity of the system, we show that co-expression in yeast of the mammalian HECT type E3 ubiquitin ligase E6AP (E6-Associated Protein) and a model substrate faithfully recapitulates E6AP-dependent substrate ubiquitination and degradation. In addition, we show that the endogenous sumoylation pathway of S. cerevisiae can specifically sumoylate mouse PML (Promyelocytic leukemia protein). In conclusion, the yeast vector system described in this paper provides a versatile tool to study complex posttranslational modifications in a cellular setting

A human neural crest model reveals the developmental impact of neuroblastoma-associated chromosomal aberrations

Early childhood tumours arise from transformed embryonic cells, which often carry large copy number alterations (CNA). However, it remains unclear how CNAs contribute to embryonic tumourigenesis due to a lack of suitable models. Here we employ female human embryonic stem cell (hESC) differentiation and single-cell transcriptome and epigenome analysis to assess the effects of chromosome 17q/1q gains, which are prevalent in the embryonal tumour neuroblastoma (NB). We show that CNAs impair the specification of trunk neural crest (NC) cells and their sympathoadrenal derivatives, the putative cells-of-origin of NB. This effect is exacerbated upon overexpression of MYCN, whose amplification co-occurs with CNAs in NB. Moreover, CNAs potentiate the pro-tumourigenic effects of MYCN and mutant NC cells resemble NB cells in tumours. These changes correlate with a stepwise aberration of developmental transcription factor networks. Together, our results sketch a mechanistic framework for the CNA-driven initiation of embryonal tumours

Devaluation of low-quality food during early experience by sheep

A ruminant's early experience with low-quality food (LQF) is expected to increase its acceptance and preference in adulthood. Contrarily, we found that experienced sheep (ES) exposed to mature oat hay early in life ate less of this LQF than inexperienced sheep (IS). A possibility is that ES might have devaluated the LQF through continuous comparisons against high-quality food (HQF) supplements (sunflower meal and ground corn) that were simultaneously available during early experience. In this study, we tested the devaluation hypothesis with a successive negative contrast (SNC) procedure. In a consummatory SNC procedure, ‘shift’ subjects are unexpectedly changed from HQF to LQF, and their consumption is then compared against the consumption of ‘unshift’ subjects that receive LQF all throughout the SNC procedure. The magnitude of the difference in consumption between preshift and postshift is regarded as a measure of the degree to which both foods (HQF and LQF) are perceived to differ hedonically. When sheep from our previous study were 300 days old, both ES and IS were randomly assigned to either shift (ES-S and IS-S) or unshift conditions (ES-U and IS-U; n = 6 in each group). Groups ES-S and IS-S were fed HQF (alfalfa hay) during the preshift phase, and then suddenly changed to LQF (oat hay) in the postshift phase. Groups ES-U and IS-U (controls) were fed only LQF throughout the SNC procedure. Subjects in ES-S showed a significantly lower intake of LQF than those in ES-U in the first postshift session (i.e. they showed an SNC effect), which was not observed in IS. These results agree with ES subjects having devalued LQF during early experience. We discuss the possibility that high levels of nutrient supplementation can result in devaluation of LQF (i.e. decrease in preference and acceptance), whereas restricted levels of supplementation may promote a positive experience with LQF.Fil: Catanese, Francisco Hernan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; ArgentinaFil: Freidin, Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; ArgentinaFil: Cuello, Marina Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; ArgentinaFil: Distel, Roberto Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; Argentin

Life in a temperate Polar sea: a unique taphonomic window on the structure of a Late Cretaceous Arctic marine ecosystem

As the earth faces a warming climate, the rock record reminds us that comparable climatic scenarios have occurred before. In the Late Cretaceous, Arctic marine organisms were not subject to frigid temperatures but still contended with seasonal extremes in photoperiod. Here, we describe an unusual fossil assemblage from Devon Island, Arctic Canada, that offers a snapshot of a ca 75 Myr ago marine palaeoecosystem adapted to such conditions. Thick siliceous biogenic sediments and glaucony sands reveal remarkably persistent high primary productivity along a high-latitude Late Cretaceous coastline. Abundant fossil faeces demonstrate that this planktonic bounty supported benthic invertebrates and large, possibly seasonal, vertebrates in short food chains. These ancient organisms filled trophic roles comparable to those of extant Arctic species, but there were fundamental differences in resource dynamics. Whereas most of the modern Arctic is oligotrophic and structured by resources from melting sea ice, we suggest that forested terrestrial landscapes helped support the ancient marine community through high levels of terrigenous organic input

Av egen kraft tillsammans med andra. Personer med psykiska funktionshinder, socialt stöd och återhämtning

The purpose of this study was to describe and analyse the significance of social support to adults with psychiatric disabilities, based on the individual’s experiences. This includes research questions concerning what type of social support was perceived to be beneficial or restrictive, which relational aspects were considered having a beneficial or restrictive effect on the support- and recovery process, as well as what strategies and capacities the individual developed on their own in order to cope with their obstacles and disadvantages. The empirical material was based on qualitative inteviews with ten individuals that defined themselves as having psychiatric disabilities. They were each interviewed on three occasions. The repetetive form of the interviews lead to three separate interview guides being created over the course of the data collection process. The goal of the first interview was to get a general idea of the subject’s daily life, activities, arenas, relationships and support. The second interview focused on meaningful relationships. The third and final interview had one general and one individual part. The general part focused on the subject’s psychiatric disabilities, their opportunities as well as existing and desired support measures. The subject matters in the individual part were related to the previous interviews with each of the participants and aimed to develop or expand on previous discussion. The material was analysed using theories about social support and recovery and was presented with roots in four different networking entities: close and extended family, friends, coworkers and fellow students, as well as formal support instances. The result showed that it is the support with mobilising qualities that is beneficial to the recovery process. It increases the individual’s degree of control over their illness, obstacles and challenges. In addition, it appeals to the individual as a central actor and as a person with difficulties as well as resources. The fact that the social support from the various support agencies have varying focus and direction benefits recovery since it increases the individual’s opportunity to maintain as well as develop their social bonds and roles within the community. A relational prerequisite for social support is an equal relationship, which benefits the recovery process as it encompasses acknowledgement and acceptance of both the individual on a personal level and of their situation. The type of social support that hinders recovery is the kind that lacks a connection between the individual’s perceived problems, their need for support and their over time fluctuating ability to handle their psychiatric disability. This is due to the fact that it robs the individual of control over the situation, increases the need for support and strengthens their perception of themselves as sick. Recovery comes to a halt when the individual isn’t included in the support and recovery process, which is clarified in relational aspects such as lacking attentiveness, subpar acceptance as well as rejection. The individual’s will and capacity to actively take on the responsibility to steer their recovery in a positive direction was made visible through four groups of strategies that were crystallised through the course of the research; creation of understanding and legitimisation, cultivation, including strategies and the creation of boundaries