Counterregulation of cAMP-directed kinase activities controls ciliogenesis

The primary cilium emanates from the cell surface of growth-arrested cells and plays a central role in vertebrate development and tissue homeostasis. The mechanisms that control ciliogenesis have been extensively explored. However, the intersection between GPCR signaling and the ubiquitin pathway in the control of cilium stability is unknown. Here, we observe that cAMP elevation promotes cilia resorption. At centriolar satellites, we identify a multimeric complex nucleated by PCM1 that includes two kinases, NEK10 and PKA, and the E3 ubiquitin ligase CHIP. We show that NEK10 is essential for ciliogenesis in mammals and for the development of medaka fish. PKA phosphorylation primes NEK10 for CHIP-mediated ubiquitination and proteolysis resulting in cilia resorption. Dearangement of this control mechanism occurs in proliferative and genetic disorders. These findings unveil a pericentriolar kinase signalosome that efficiently links the cAMP cascade with the ubiquitin-proteasome system, controlling essential aspects of ciliogenesis

praja2 regulates KSR1 stability and mitogenic signaling

The kinase suppressor of Ras 1 (KSR1) has a fundamental role in mitogenic signaling by scaffolding components of the Ras/MAP kinase pathway. In response to Ras activation, KSR1 assembles a tripartite kinase complex that optimally transfers signals generated at the cell membrane to activate ERK. We describe a novel mechanism of ERK attenuation based on ubiquitin-dependent proteolysis of KSR1. Stimulation of membrane receptors by hormones or growth factors induced KSR1 polyubiquitination, which paralleled a decline of ERK1/2 signaling. We identified praja2 as the E3 ligase that ubiquitylates KSR1. We showed that praja2-dependent regulation of KSR1 is involved in the growth of cancer cells and in the maintenance of undifferentiated pluripotent state in mouse embryonic stem cells. The dynamic interplay between the ubiquitin system and the kinase scaffold of the Ras pathway shapes the activation profile of the mitogenic cascade. By controlling KSR1 levels, praja2 directly affects compartmentalized ERK activities, impacting on physiological events required for cell proliferation and maintenance of embryonic stem cell pluripotency

Restoration of CFTR function in patients with cystic fibrosis carrying the F508del-CFTR mutation

<div><p>Restoration of BECN1/Beclin 1-dependent autophagy and depletion of SQSTM1/p62 by genetic manipulation or autophagy-stimulatory proteostasis regulators, such as cystamine, have positive effects on mouse models of human cystic fibrosis (CF). These measures rescue the functional expression of the most frequent pathogenic CFTR mutant, F508del, at the respiratory epithelial surface and reduce lung inflammation in <i>Cftr^F508del</i> homozygous mice. Cysteamine, the reduced form of cystamine, is an FDA-approved drug. Here, we report that oral treatment with cysteamine greatly reduces the mortality rate and improves the phenotype of newborn mice bearing the <i>F508del-CFTR</i> mutation. Cysteamine was also able to increase the plasma membrane expression of the F508del-CFTR protein in nasal epithelial cells from <i>F508del</i> homozygous CF patients, and these effects persisted for 24 h after cysteamine withdrawal. Importantly, this cysteamine effect after washout was further sustained by the sequential administration of epigallocatechin gallate (EGCG), a green tea flavonoid, both <i>in vivo</i>, in mice, and <i>in vitro</i>, in primary epithelial cells from CF patients. In a pilot clinical trial involving 10 <i>F508del-CFTR</i> homozygous CF patients, the combination of cysteamine and EGCG restored BECN1, reduced SQSTM1 levels and improved CFTR function from nasal epithelial cells <i>in vivo</i>, correlating with a decrease of chloride concentrations in sweat, as well as with a reduction of the abundance of <i>TNF/TNF-alpha (tumor necrosis factor)</i> and <i>CXCL8</i> (<i>chemokine [C-X-C motif] ligand 8</i>) transcripts in nasal brushing and TNF and CXCL8 protein levels in the sputum. Altogether, these results suggest that optimal schedules of cysteamine plus EGCG might be used for the treatment of CF caused by the <i>F508del-CFTR</i> mutation.</p></div

TRAP1-dependent regulation of p70S6K is involved in the attenuation of protein synthesis and cell migration: Relevance in human colorectal tumors

TNF receptor-associated protein 1 (TRAP1) is an HSP90 chaperone involved in stress protection and apoptosis in mitochondrial and extramitochondrial compartments. Remarkably, aberrant deregulation of TRAP1 function has been observed in several cancer types with potential new opportunities for therapeutic intervention in humans. Although previous studies by our group identified novel roles of TRAP1 in quality control of mitochondria-destined proteins through the attenuation of protein synthesis, molecular mechanisms are still largely unknown. To shed further light on the signaling pathways regulated by TRAP1 in the attenuation of protein synthesis, this study demonstrates that the entire pathway of cap-mediated translation is activated in cells following TRAP1 interference: consistently, expression and consequent phosphorylation of p70S6K and RSK1, two translation activating kinases, are increased upon TRAP1 silencing. Furthermore, we show that these regulatory functions affect the response to translational stress and cell migration in wound healing assays, processes involving both kinases. Notably, the regulatory mechanisms controlled by TRAP1 are conserved in colorectal cancer tissues, since an inverse correlation between TRAP1 and p70S6K expression is found in tumor tissues, thereby supporting the relevant role of TRAP1 translational regulation in vivo. Taken as a whole, these new findings candidate TRAP1 network for new anti-cancer strategies aimed at targeting the translational/quality control machinery of tumor cells

Parietal maps of visual signals for bodily action planning

The posterior parietal cortex (PPC) has long been understood as a high-level integrative station for computing motor commands for the body based on sensory (i.e., mostly tactile and visual) input from the outside world. In the last decade, accumulating evidence has shown that the parietal areas not only extract the pragmatic features of manipulable objects, but also subserve sensorimotor processing of others’ actions. A paradigmatic case is that of the anterior intraparietal area (AIP), which encodes the identity of observed manipulative actions that afford potential motor actions the observer could perform in response to them. On these bases, we propose an AIP manipulative action-based template of the general planning functions of the PPC and review existing evidence supporting the extension of this model to other PPC regions and to a wider set of actions: defensive and locomotor actions. In our model, a hallmark of PPC functioning is the processing of information about the physical and social world to encode potential bodily actions appropriate for the current context. We further extend the model to actions performed with man-made objects (e.g., tools) and artifacts, because they become integral parts of the subject’s body schema and motor repertoire. Finally, we conclude that existing evidence supports a generally conserved neural circuitry that transforms integrated sensory signals into the variety of bodily actions that primates are capable of preparing and performing to interact with their physical and social world

3D strip model for continuous roll-forming process simulation

Abstract The paper addresses the complexities for a reliable numerical simulation of the roll forming process. During the process, the material is progressively bent accumulating plastic deformation at each forming step. Strain hardening limits the material formability and may causes flaws of the final shape. A simplified method for the FEM modeling of the process has been developed introducing a narrow-strip 3D model. This approach leads better performance than the classical modeling method, in terms of results reliability and low computational time. In order to verify the proposed model, an experimental campaign of testing, for a specific roll forming production process, was carried out. On the quasi-static regime, the post necking behavior of the sheet metal was characterized. The Vickers hardness and the plastic strain of uniaxial tests were empirically correlated. By the hardness correlation, the plastic strain accumulated at different stages of the process was evaluated and compared with the numerical results. Further possible improvements of the method are highlighted

Global Study: Participation in One Health Networks and Involvement in COVID-19 Response Activities

PURPOSE: This global study examined whether being part of a One Health Network (OHN) was associated with being involved in COVID-19 response activities at the early stages of the pandemic. Barriers to workforce involvement in the pandemic response and the perceived value of OHN activities were studied to inform future targeted evidence-based strategies for workforce capacity-building. METHODS & MATERIALS: We conducted a cross-sectional descriptive study, using an online questionnaire that was globally distributed in July-August 2020. With a snowball sampling approach via OHN listservs, social media, and further sharing, we aimed to reach individuals in the global health workforce across locations, organizations, and sectors to survey their participation in OHN activities and involvement in COVID-19 response. RESULTS: The sample included 1050 respondents from various types of organizations and work sectors, from 94 countries across all WHO regions. Being part of an OHN was positively associated with involvement in the COVID-19 response (odds ratio: 1.8, 95% confidence interval: 1.3 - 2.4). The OHN activities most indicated as useful during COVID-19 pandemic by the survey respondents included 'increased public awareness of One Health' and 'networking with professionals across sectors with common interests'. Overall, 44% of survey respondents who were part of an OHN found OHN activities very or extremely helpful to their COVID-19 response. Lack of opportunities was a commonly reported barrier to involvement in COVID-19 response globally, and lack of funding was a barrier particularly in the WHO Africa region. CONCLUSION: This study provides a snapshot of the multisectoral response to COVID-19 and an assessment of the contribution of OHNs. The lessons learned during this pandemic can be used to identify measures to improve global health capacity, including OHN activities to build and strengthen workforce response to future global health challenges