Targeting of the prion protein to the cytosol: mechanisms and consequences

Prion diseases are characterized by the conformational transition of the cellular prion protein (PrPC) into an aberrant protein conformer, designated scrapie-prion protein (PrPSc). A causal link between protein misfolding and neurodegeneration has been established for a variety of neurodegenerative disease, such as Alzheimer's disease, Parkinson's disease and polyglutamine diseases, but there is an ongoing debate about the nature of the neurotoxic species and how non-native conformers can damage neuronal populations. PrP is normally imported into the endoplasmic reticulum (ER) and targeted to the outer leaflet of the plasma membrane via a glycosylphosphatidylinositol (GPI) anchor. However, several conditions, such as ER stress or some pathogenic mutations in the PrP gene, can induce the mislocalization of PrP in the cytosol, where it has a neurotoxic potential as demonstrated in cell culture and transgenic mouse models. In this review we focus on intrinsic factors and cellular pathways implicated in the import of PrP into the ER and its mistargeting to the cytosol. The findings summarized here not only reveal a complex regulation of the biogenesis of PrP, but also provide interesting new insight into toxic activities of pathogenic protein conformers and quality control pathways of ER-targeted proteins

Promoting Social Participation, Leisure and Community Integration for Adults with Developmental Disabilities: An Intervention Guide

Upon an initial review of literature, it was found that life expectancy of individuals with developmental disabilities has been increasing which is leading to a greater number of adults with developmental disabilities living in the community. Because these individuals are living longer, it puts an extended burden on caregivers and the adults with developmental disabilities are being placed in group homes. Adults with developmental disabilities have limited access to activities outside of group homes (Mansell, Elliott, Beadle-Brown, Ashman, & Macdonald, 2002; Abbott & McConkey, 2006). They are spending most of their time engaged in more passive activities and have little social interaction with people other than parents, staff, or other adults with developmental disabilities (Felce & Perry, 1995; & Lippold & Burns, 2009). Because of this, opportunities for social participation and leisure activities within the community are limited leading to a lack of community integration. Occupational therapists can play a vital role in the development of these skills and extend their performance range in order to engage adults with developmental disabilities into leisure activities and integrate them into the community. The product of this scholarly project, which is intended for use by occupational therapists, is an intervention guide focusing on three areas: social participation, leisure exploration and participation, and community integration for adults with developmental disabilities. An in-service presentation was also created to educate administrative personnel and staff about the issues and to advocate for use of the activities in the intervention guide group homes. The methodology for the project consisted of an extensive review of literature and resources. The interventions in the guide include those created by the authors and others that were adapted using materials from other resources and authors. Cole’s Seven Steps were not used in their entirety but were used to guide the structure of the group activities, and the Ecology of Human Performance model was foundational in the development of the product as well (Schwartzberg, Howe, & Barnes, 2009) The recommendations for this project include: further researching on available programs addressing all areas of occupation in group home settings, completing outcome research on the effectiveness of the interventions included in the guide, expanding the use of the guide to those who reside at home with caregivers, and an addition of more interventions into the guide using evidence-based practice. The limitations of this project are that it has not been implemented into practice and does not address all areas of occupation

Translokation des Prion-Proteins ins endoplasmatische Retikulum

Vorangegangene Studien zeigten, dass das Prion-Protein (PrP) an der ER-Membran in verschiedenen topologischen Isoformen synthetisiert wird und teilweise sogar im Zytosol vorliegen kann. Sowohl die ER-Signalsequenz als auch die hydrophobe Domäne von PrP wurden dabei als Domänen identifiziert, die eine Rolle in der Translokation spielen. Die hier durchgeführte Analyse des ER-Imports von PrP und verschiedenen chimären Proteinen hat nun erstmals gezeigt, dass auch der Faltungszustand von Polypeptiden Einfluss auf deren Translokation ins ER-Lumen haben kann. Die vorliegende Studie ergab, dass - unabhängig von der Primärstruktur - ein gewisses Maß an α-helikalen Bereichen notwendig für einen produktiven ER-Import ist. Sowohl die Länge der Polypeptide als auch posttranslationelle Modifikationen wie die GPI-Verankerung, die N-Glykosylierung oder die Ausbildung einer Disulfidbrücke beeinflussen die Translokation nicht. Darüber hinaus deuten die Ergebnisse der vorliegenden Arbeit darauf hin, dass Proteine mit ausgedehnten unstrukturierten Domänen am N-Terminus einer kotranslokationalen Qualitätskontrolle unterliegen und noch vor der Translokation ins ER-Lumen einer proteasomalen Degradierung im Zytosol zugeführt werden. Die in dieser Doktorarbeit dargestellten Ergebnisse legen daher die Vermutung nahe, dass die Ausbildung von Sekundärstrukturen vor oder während der Translokation die weitere Biogenese des naszierenden Polypeptids beeinflusst. Die gewonnenen Erkenntnisse können dazu beitragen die physiologischen aber auch die möglichen pathophysiologischen Konsequenzen der Regulation der Translokation besser zu verstehen. Der zweite Teil der Arbeit erbrachte erstmals experimentelle Evidenzen, dass trotz sehr geringer Sequenzhomologie zwischen den PrP-Homologen im Zebrabärbling (Danio rerio) und Säugetier-PrP die charakteristischen posttranslationalen Modifikationen, wie beispielsweise die komplexe Glykosylierung und der C-terminale GPI-Anker, konserviert sind. Die neu etablierten Zellkulturmodelle zur Analyse von PrP-homologen Proteinen deuten auf eine evolutionär konservierte Funktion von PrP hin und könnten dazu beitragen, neue Einsichten in die physiologische Aktivität von PrP zu gewinnen

Translokation des Prion-Proteins ins endoplasmatische Retikulum

Vorangegangene Studien zeigten, dass das Prion-Protein (PrP) an der ER-Membran in verschiedenen topologischen Isoformen synthetisiert wird und teilweise sogar im Zytosol vorliegen kann. Sowohl die ER-Signalsequenz als auch die hydrophobe Domäne von PrP wurden dabei als Domänen identifiziert, die eine Rolle in der Translokation spielen. Die hier durchgeführte Analyse des ER-Imports von PrP und verschiedenen chimären Proteinen hat nun erstmals gezeigt, dass auch der Faltungszustand von Polypeptiden Einfluss auf deren Translokation ins ER-Lumen haben kann. Die vorliegende Studie ergab, dass - unabhängig von der Primärstruktur - ein gewisses Maß an α-helikalen Bereichen notwendig für einen produktiven ER-Import ist. Sowohl die Länge der Polypeptide als auch posttranslationelle Modifikationen wie die GPI-Verankerung, die N-Glykosylierung oder die Ausbildung einer Disulfidbrücke beeinflussen die Translokation nicht. Darüber hinaus deuten die Ergebnisse der vorliegenden Arbeit darauf hin, dass Proteine mit ausgedehnten unstrukturierten Domänen am N-Terminus einer kotranslokationalen Qualitätskontrolle unterliegen und noch vor der Translokation ins ER-Lumen einer proteasomalen Degradierung im Zytosol zugeführt werden. Die in dieser Doktorarbeit dargestellten Ergebnisse legen daher die Vermutung nahe, dass die Ausbildung von Sekundärstrukturen vor oder während der Translokation die weitere Biogenese des naszierenden Polypeptids beeinflusst. Die gewonnenen Erkenntnisse können dazu beitragen die physiologischen aber auch die möglichen pathophysiologischen Konsequenzen der Regulation der Translokation besser zu verstehen. Der zweite Teil der Arbeit erbrachte erstmals experimentelle Evidenzen, dass trotz sehr geringer Sequenzhomologie zwischen den PrP-Homologen im Zebrabärbling (Danio rerio) und Säugetier-PrP die charakteristischen posttranslationalen Modifikationen, wie beispielsweise die komplexe Glykosylierung und der C-terminale GPI-Anker, konserviert sind. Die neu etablierten Zellkulturmodelle zur Analyse von PrP-homologen Proteinen deuten auf eine evolutionär konservierte Funktion von PrP hin und könnten dazu beitragen, neue Einsichten in die physiologische Aktivität von PrP zu gewinnen

Association of Bcl-2 with misfolded prion protein is linked to the toxic potential of cytosolic PrP

Protein misfolding is linked to different neurodegenerative disorders like Alzheimer’s disease, polyglutamine, and prion diseases. We investigated the cytotoxic effects of aberrant conformers of the prion protein (PrP) and show that toxicity is specifically linked to misfolding of PrP in the cytosolic compartment and involves binding of PrP to the anti-apoptotic protein Bcl-2. PrP targeted to different cellular compartments, including the cytosol, nucleus, and mitochondria, adopted a misfolded and partially proteinase K–resistant conformation. However, only in the cytosol did the accumulation of misfolded PrP induce apoptosis. Apoptotic cell death was also induced by two pathogenic mutants of PrP, which are partially localized in the cytosol. A mechanistic analysis revealed that the toxic potential is linked to an internal domain of PrP (amino acids 115–156) and involves coaggregation of cytosolic PrP with Bcl-2. Increased expression of the chaperones Hsp70 and Hsp40 prevented the formation of PrP/Bcl-2 coaggregates and interfered with PrP-induced apoptosis. Our study reveals a compartment-specific toxicity of PrP misfolding that involves coaggregation of Bcl-2 and indicates a protective role of molecular chaperones

α-Helical Domains Promote Translocation of Intrinsically Disordered Polypeptides into the Endoplasmic Reticulum

Co-translational import into the endoplasmic reticulum (ER) is primarily controlled by N-terminal signal sequences that mediate targeting of the ribosome-nascent chain complex to the Sec61/translocon and initiate the translocation process. Here we show that after targeting to the translocon the secondary structure of the nascent polypeptide chain can significantly modulate translocation efficiency. ER-targeted polypeptides dominated by unstructured domains failed to efficiently translocate into the ER lumen and were subjected to proteasomal degradation via a co-translocational/preemptive pathway. Productive ER import could be reinstated by increasing the amount of α-helical domains, whereas more effective ER signal sequences had only a minor effect on ER import efficiency of unstructured polypeptides. ER stress and overexpression of p58IPK promoted the co-translocational degradation pathway. Moreover polypeptides with unstructured domains at their N terminus were specifically targeted to proteasomal degradation under these conditions. Our study indicates that extended unstructured domains are signals to dispose ER-targeted proteins via a co-translocational, preemptive quality control pathway

Annex 65, Long-Term Performance of Super-Insulating-Materials in Building Components and Systems. Report of Subtask II: Scientific Information for Standardization Bodies dealing with Hygro-Thermo-Mechanical Properties and Ageing

This subtask is divided in two actions:Action 2A: Materials Assessment & Ageing Procedures (Experiments & Simulation)Action 2B: Components & Systems Assessment (Experiments & Simulation)As their structures and microstructures are completely different, Super-Insulating Materials (SIMs) cannot be compared directly to traditional insulating materials. Worldwide acceptance of these materials will be improved if the hygro-thermal and mechanical properties of SIM can be clearly articulated and reproduced. In particular, nano-structured materials used to manufacture a SIM are characterized by a high specific area (m\ub2/g) and narrow pores (smaller than 1 μm) which make them very sensitive to gas adsorption and condensation, especially in contact with water molecules.Therefore, methods of characterization must be adapted, or new methods developed to measure the microstructural, hygro-thermal and mechanical properties of these materials and their barrier films.In parallel, modelling methods to describe heat, moisture and air transfer through nano-structured materials and films will have to be developed (adsorption and desorption models, diffusion models, freeze-thawing …).Of course, a few methods will be common to all SIMs, but due to their structural differences some specific modelling methods have to be developed.SIMs can offer considerable advantages (low thickness, low Uvalue) ; however potential drawback effects should be considered in the planning process in order to optimise the development of these extraordinary properties (very low thermal conductivity) and to prevent negative publicity which could be detrimental to this sector of emerging products. This is why ageing tests will be set according to realistic conditions (temperature, moisture, pressure, load …) as set out in SubTask 3A. One objective of artificial ageing is to understand potential degradation processes that could occur. The durability of hydrophobic treatment is one of these processes and will also be subject to discussion and investigation.At the component scale, additional characterizations are needed as panels or rolls are sold by manufacturers. In particular, thermal bridges will be carefully investigated, as the extraordinary thermal performance of SIMs are sensitive to the influence of thermal bridges

Using botanic gardens and arboreta to help identify urban trees for the future

Societal Impact StatementDiversification of urban forests is essential to enhance their resilience to future biotic threats as well as those posed by a changing climate. Arboreta and botanic gardens host a wide range of plant material that can be evaluated to inform tree selection policy. This study demonstrates that plant functional traits, such as the water potential at leaf turgor loss, can be highly instructive when developing evidence-based recommendations for urban environments. However, if botanic collections are to fulfil a critical role in understanding plant response to environment, they should not be managed solely as visitor attractions but must have scientific objectives at the forefront of management policy.SummaryArboreta and botanic gardens host a multitude of species that can be utilized in research focused on improving diversity within urban forests. Higher tree species diversity will enhance the resilience of urban forests to abiotic and biotic threats and help deliver strategies that foster sustainable communities. Consequently, this study aims to demonstrate the value of botanic collections as a resource for research into tree species selection for more resilient urban landscapes. As water stress is a major constraint for trees in urban environments, understanding the drought tolerance of species is essential for urban tree selection. This study evaluates a key functional trait relating to drought tolerance. Using vapor pressure osmometry, the water potential at leaf turgor loss was evaluated for 96 species using plant material from seven botanic collections in North America and Europe. Leaf turgor loss contrasted widely in the temperate deciduous trees evaluated and, in summer, ranged from -1.7 MPa to -3.9 MPa. Significant differences in drought tolerance were also apparent across genera and closely related cultivars. Osmotic adjustment was shown to be a major physiological factor driving leaf turgor loss. A meta-analysis also demonstrated that leaf turgor loss was closely related to a drought-tolerance scale based on observations of tree performance under drought. Arboreta and botanic collections can play a vital role in the evaluation of plant material for urban environments, provided they are curated with scientific objectives at the forefront of management policy and are not managed purely as visitor attractions

Anti-prion drug mPPIg5 inhibits PrP(C) conversion to PrP(Sc).

Prion diseases, also known as transmissible spongiform encephalopathies, are a group of fatal neurodegenerative diseases that include scrapie in sheep, bovine spongiform encephalopathy (BSE) in cattle and Creutzfeldt-Jakob disease (CJD) in humans. The 'protein only hypothesis' advocates that PrP(Sc), an abnormal isoform of the cellular protein PrP(C), is the main and possibly sole component of prion infectious agents. Currently, no effective therapy exists for these diseases at the symptomatic phase for either humans or animals, though a number of compounds have demonstrated the ability to eliminate PrPSc in cell culture models. Of particular interest are synthetic polymers known as dendrimers which possess the unique ability to eliminate PrP(Sc) in both an intracellular and in vitro setting. The efficacy and mode of action of the novel anti-prion dendrimer mPPIg5 was investigated through the creation of a number of innovative bio-assays based upon the scrapie cell assay. These assays were used to demonstrate that mPPIg5 is a highly effective anti-prion drug which acts, at least in part, through the inhibition of PrP(C) to PrP(Sc) conversion. Understanding how a drug works is a vital component in maximising its performance. By establishing the efficacy and method of action of mPPIg5, this study will help determine which drugs are most likely to enhance this effect and also aid the design of dendrimers with anti-prion capabilities for the future

Predictors of tropical cyclone-induced urban tree failure: an international scoping review

Background: Trees are critical components of rural and urban ecosystems throughout the world. While they have adapted to the historic conditions of their native environments, climate change, urbanization, and human-assisted range expansion may test the storm resiliency of many tree species. Objective: In this global multilingual scoping review, we investigate a range of intrinsic (i.e., tree characteristics) and external (i.e., environmental and management) factors which have been used to predict tree failure during tropical cyclones. Design: We searched online databases and journals in English, Chinese, French, Japanese, Portuguese, and Spanish to find peer-reviewed papers and dissertations. We retained papers that used ground-based methods to study tree damage following a tropical cyclone and conducted a statistical analysis of factors that influence tree resistance to damage. From each paper we extracted details of study methods, and the relationships between damage and predictors. Results: Our efforts generated 65 peer-reviewed papers and dissertations that met our final criteria for inclusion (i.e., data on the relative proportion of trees failed/intact as assessed no more than a year after the storm event). Of these papers 37 independent variables were assessed to predict tree failure. Research in both urban and rural settings tends to be concentrated in regions frequently impacted by tropical cyclones. Characteristics of species such as wood density have been studied in rural environments and are also relevant predictors for tree failure in urban trees. Environmental characteristics unique to urban settings such as planting areas surrounded by pavement need further research. Several urban studies demonstrate that risk assessment methods can predict tree failure during a storm. Conclusion: Results can be used by future storm researchers to identify both predictors may warrant inclusion in their models as well as predictors which have yet to be tested. Results can also inform planning and activities that can mitigate tropical cyclone damage to the urban forest