Negative Regulator of Ubiquitin-Like Protein 1 modulates the autophagy-lysosomal pathway via p62 to facilitate the extracellular release of tau following proteasome impairment

Negative regulator of ubiquitin-like protein 1 (NUB1) and its longer isoform NUB1L are ubiquitin-like (UBL)/ubiquitin-associated (UBA) proteins that facilitate the targeting of proteasomal substrates, including tau, synphilin-1 and huntingtin. Previous data revealed that NUB1 also mediated a reduction in tau phosphorylation and aggregation following proteasome inhibition, suggesting a switch in NUB1 function from targeted proteasomal degradation to a role in autophagy. Here, we delineate the mechanisms of this switch and show that NUB1 interacted specifically with p62 and induced an increase in p62 levels in a manner facilitated by inhibition of the proteasome. NUB1 moreover increased autophagosomes and the recruitment of lysosomes to aggresomes following proteasome inhibition. Autophagy flux assays revealed that NUB1 affected the autophagy–lysosomal pathway primarily via the UBA domain. NUB1 localized to cytosolic inclusions with pathological forms of tau, as well as LAMP1 and p62 in the hippocampal neurons of tauopathy mice. Finally, NUB1 facilitated the extracellular release of tau following proteasome inhibition. This study thus shows that NUB1 plays a role in regulating the autophagy–lysosomal pathway when the ubiquitin proteasome system is compromised, thus contributing to the mechanisms targeting the removal of aggregation-prone proteins upon proteasomal impairment

Functional expression of TMEM16A in taste bud cells

Key points: Taste transduction occurs in taste buds in the tongue epithelium. The Ca2+-activated Cl– channels TMEM16A and TMEM16B play relevant physiological roles in several sensory systems. Here, we report that TMEM16A, but not TMEM16B, is expressed in the apical part of taste buds. Large Ca2+-activated Cl− currents blocked by Ani-9, a selective inhibitor of TMEM16A, are measured in type I taste cells but not in type II or III taste cells. ATP indirectly activates Ca2+-activated Cl– currents in type I cells through TMEM16A channels. These results indicate that TMEM16A is functional in type I taste cells and contribute to understanding the largely unknown physiological roles of these cells. Abstract: The Ca2+-activated Cl– channels TMEM16A and TMEM16B have relevant roles in many physiological processes including neuronal excitability and regulation of Cl– homeostasis. Here, we examined the presence of Ca2+-activated Cl– channels in taste cells of mouse vallate papillae by using immunohistochemistry and electrophysiological recordings. By using immunohistochemistry we showed that only TMEM16A, and not TMEM16B, was expressed in taste bud cells where it largely co-localized with the inwardly rectifying K+ channel KNCJ1 in the apical part of type I cells. By using whole-cell patch-clamp recordings in isolated cells from taste buds, we measured an average current of −1083 pA at −100 mV in 1.5 μm Ca2+ and symmetrical Cl– in type I cells. Ion substitution experiments and blockage by Ani-9, a specific TMEM16A channel blocker, indicated that Ca2+ activated anionic currents through TMEM16A channels. We did not detect any Ca2+-activated Cl– currents in type II or III taste cells. ATP is released by type II cells in response to various tastants and reaches type I cells where it is hydrolysed by ecto-ATPases. Type I cells also express P2Y purinergic receptors and stimulation of type I cells with extracellular ATP produced large Ca2+-activated Cl− currents blocked by Ani-9, indicating a possible role of TMEM16A in ATP-mediated signalling. These results provide a definitive demonstration that TMEM16A-mediated currents are functional in type I taste cells and provide a foundation for future studies investigating physiological roles for these often-neglected taste cells

Allele-specific editing ameliorates dominant retinitis pigmentosa in a transgenic mouse model

Retinitis pigmentosa (RP) is a group of progressive retinal degenerations of mostly monogenic inheritance, which cause blindness in about 1:3,500 individuals worldwide. Heterozygous variants in the rhodopsin (RHO) gene are the most common cause of autosomal dominant RP (adRP). Among these, missense variants at C-terminal proline 347, such as p.Pro347Ser, cause severe adRP recurrently in European affected individuals. Here, for the first time, we use CRISPR/Cas9 to selectively target the p.Pro347Ser variant while preserving the wild-type RHO allele in vitro and in a mouse model of adRP. Detailed in vitro, genomic, and biochemical characterization of the rhodopsin C-terminal editing demonstrates a safe downregulation of p.Pro347Ser expression leading to partial recovery of photoreceptor function in a transgenic mouse model treated with adeno-associated viral vectors. This study supports the safety and efficacy of CRISPR/Cas9-mediated allele-specific editing and paves the way for a permanent and precise correction of heterozygous variants in dominantly inherited retinal diseases

Productive integration, economic recession and employment in Europe: an assessment based on vertically integrated sectors

The Covid-19 crisis has revamped the discussion about the redefinition of GVC. This paper contributes to the debate, analysing the productive relationships between European countries in four key manufacturing activities. In particular, the paper addresses two objectives. First, it maps the degree of productive integration in Europe, focusing on the generation of employment in the production of exported intermediate inputs and final goods. Second, it provides a preliminary assessment of the potential impact on employment that the current economic crisis will have on some manufacturing activities across Europe. The analysis is realised employing the concept of vertically integrated labour (Pasinetti 1973) which allows to account for the employment directly and indirectly involved in the production of final goods. The estimations are derived from Multi-Regional Input–Output tables to map the supply chain and to differentiate between the employment involved in the production of exported intermediate inputs and final goods. The results show that most of the employment involved in the production of final output of the activities studied in the paper is linked to international trade. Although Europe shows a high degree of productive links, there are important differences in the modality of insertion in the productive structure of European countries. Moreover, the impact on the level of employment due to the current economic crisis can be significant, affecting more than 1.3 million of people in Europe. These results are relevant to policy makers, who should consider carefully the high degree of linkages of the European economies when designing industrial policies and measure of support to the economy

Did Italy Need More Labour Flexibility?: The Consequences of the Jobs Act

The dramatic impact of the 2008 crisis on the Italian economy led to policy responses including structural reforms and labour market liberalisation to reverse the worrisome output and employment trends. A key action by the Italian government, the evocatively named Jobs Act of 2014, has deeply changed Italian industrial relations. The Jobs Act has introduced a new contract type that substantially limits workers’ rights to reinstatement in case of fi rms invalidly fi ring them. This article frames the Jobs Act within the overall liberalisation process begun in Italy in the 1990s, providing an initial evaluation of its impacts. Using detailed data sources, we show that the expected boost in employment cannot be detected, the share of temporary contracts over open-ended ones has increased and the number of part-time contracts has risen. This evidence suggests that the Jobs Act is failing to achieve its main goals

Skill mismatch and the dynamics of Italian companies’ productivity

This work explores the relationship between labour productivity and skill (mis) match relying on a unique database integrating information at both the firm and the worker level. The analysis is based on a novel skill match indicator providing actual and qualitatively detailed information on the demand/supply of skills. Focusing on a sample of Italian limited liability companies observed during 2012, 2014 and 2017, we show that the ability to match their skills need via new hires is always positively correlated to companies’ labour productivity. This result is robust to the inclusion of variables accounting for sectoral-level training intensity, firm-level recruitment behaviour, a capillary set of firm-level controls and across size classes

Working from home and the explosion of enduring divides: income, employment and safety risks

Why are there so many non-teleworkable occupations? Is teleworking only a matter of ICT usage or does it also reflect the division of labour and the underlying hierarchical layers inside organizations? What does it happen to those workers not able to telework in terms of socio-economic risks, and how does the gender dimension interact with risk stratification? Hereby, we intend to shed light on these questions using a detailed integrated dataset at individual and occupational level (Indagine Campionaria delle Professioni, Indagine delle Forze di Lavoro and Inail archive) which provides information on different nature of risks (income, employment and safety). Our results entail that, first, class attributes, intended as execution of tasks, degrees of autonomy in doing the job, layers of the occupational categories, strongly influence the chance of working from home; second, those individuals who are not able to perform their work remotely are more exposed to transition to unemployment, to earn low wages, and to safety and health risks; third, being woman and employed with a temporary contract significantly amplify risk stratification