A reversible phospho-switch mediated by ULK1 regulates the activity of autophagy protease ATG4B

Upon induction of autophagy, the ubiquitin-like protein LC3 is conjugated to phosphatidylethanolamine (PE) on the inner and outer membrane of autophagosomes to allow cargo selection and autophagosome formation. LC3 undergoes two processing steps, the proteolytic cleavage of pro-LC3 and the de-lipidation of LC3-PE from autophagosomes, both executed by the same cysteine protease ATG4. How ATG4 activity is regulated to co-ordinate these events is currently unknown. Here we find that ULK1, a protein kinase activated at the autophagosome formation site, phosphorylates human ATG4B on serine 316. Phosphorylation at this residue results in inhibition of its catalytic activity in vitro and in vivo. On the other hand, phosphatase PP2A-PP2R3B can remove this inhibitory phosphorylation. We propose that the opposing activities of ULK1-mediated phosphorylation and PP2A-mediated dephosphorylation provide a phospho-switch that regulates the cellular activity of ATG4B to control LC3 processing

Data from a multidisciplinary poll of 178 expert physicians on the usage of non-vitamin K Oral Anticoagulants in patients with atrial fibrillation and venous thromboembolism

This data article contains data from a multidisciplinary questionnaires filled in by 178 expert physicians on the usage of non-vitamin K Oral Anticoagulants in patients with atrial fibrillation (AF) and for the treatment of patients with venous thromboembolism (VTE). The questionnaire consists of 9 statements of clinical complex AF and VTE cases and informative campaign on antithrombotic therapy for stroke prevention in AF. The data are potentially valuable for the scientific community, showing the doubts of different specialists (Internists, Pneumologists, Geriatricians, Cardiologists and Neurologists) with a large experience in prescribing oral anticoagulation in difficult AF and VTE cases (see full list of participants provided). The data obtained in some particular clinical cases such as CHA2DS2-VASc=1, comorbid coronary artery disease, frailty, advanced age, risk of falling and prior haemorrhagic stroke, can be compared with indications from published guidelines and recommendations for future insight and to be considered as a benchmark for future trials in the area or oral anticoagulation for AF and VTE.The data concerning informative campaign on antithrombotic therapy for stroke prevention showed the expert panel agreement on the inclusion of self monitoring of heart rhythm by pulse taking in subjects older than 64 years of age (81% agreement, item 3); knowledge that the risk of stroke associated with AF is almost twice the risk associated with hypertension (95% agreement, item 4); knowledge that the CHA2DS2-VASc score exerts a higher influence on stroke risk compared to AF duration (92% agreement, item 5); knowledge that stroke prevention in AF with a NOAC is more effective, does not cause any higher bleeding risk, and is equally simple compared to aspirin treatment (91% agreement, item 6).Data on strategies to optimise appropriate prescription of antithrombotic therapy showed agreement on the utility of short television advertisements about the risks of stroke associated with AF (79% agreement, item 8), on a campaign encouraging regular control of cardiac rhythm by pulse taking (77% agreement, item 1), on a campaign reporting the advantages of anticoagulation over no antithrombotic therapy (98% agreement, item 2) or of NOACs over aspirin (96% agreement, item 3) or on the practical use of NOAC (93% agreement, item 6) or on stroke and bleeding risk scores (87% agreement, item 7). See Colonna et al. (2017) [1] for further interpretation and discussion

Axion search with a quantum-limited ferromagnetic haloscope

A ferromagnetic axion haloscope searches for Dark Matter in the form of axions by exploiting their interaction with electronic spins. It is composed of an axion-to-electromagnetic field transducer coupled to a sensitive rf detector. The former is a photon-magnon hybrid system, and the latter is based on a quantum-limited Josephson parametric amplifier. The hybrid system consists of ten 2.1 mm diameter YIG spheres coupled to a single microwave cavity mode by means of a static magnetic field. Our setup is the most sensitive rf spin-magnetometer ever realized. The minimum detectable field is $5.5\times10^{-19}\,$T with 9 h integration time, corresponding to a limit on the axion-electron coupling constant $g_{aee}\le1.7\times10^{-11}$ at 95% CL. The scientific run of our haloscope resulted in the best limit on DM-axions to electron coupling constant in a frequency span of about 120 MHz, corresponding to the axion mass range $42.4$-$43.1\,\mu$eV. This is also the first apparatus to perform an axion mass scanning by changing the static magnetic field.Comment: 4 pages, 4 figure

Cryogenic Characteristics of the ATLAS Barrel Toroid Superconducting Magnet

ATLAS, one of the experiments of the LHC accelerator under commissioning at CERN, is equipped with a large superconducting magnet the Barrel Toroid (BT) that has been tested at nominal current (20500 A). The BT is composed of eight race-track superconducting coils (each one weights about 45 tons) forming the biggest air core toroidal magnet ever built. By means of a large throughput centrifugal pump, a forced flow (about 10 liter/second at 4.5 K) provides the indirect cooling of the coils in parallel. The paper describes the results of the measurements carried out on the complete cryogenic system assembled in the ATLAS cavern situated 100 m below the ground level. The measurements include, among other ones, the static heat loads, i.e., with no or constant current in the magnet, and the dynamic ones, since additional heat losses are produced, during the current ramp-up or slow dump, by eddy currents induced on the coil casing

Redundancy of human ATG4 protease isoforms in autophagy and LC3/GABARAP processing revealed in cells

Macroautophagy/autophagy is a cellular degradation pathway that delivers cytoplasmic material to lysosomes via double-membrane organelles called autophagosomes. Lipidation of ubiquitin-like LC3/GABARAP proteins on the autophagosome membrane is important for autophagy. The cysteine protease ATG4 executes 2 LC3/GABARAP processing events: priming of newly synthesized pro-LC3/GABARAP to enable subsequent lipidation, and delipidation/deconjugation of lipidated LC3/GABARAP (the exact purpose of which is unclear in mammals). Four ATG4 isoforms (ATG4A to ATG4D) exist in mammals; however, the functional redundancy of these proteins in cells is poorly understood. Here we show that human HAP1 and HeLa cells lacking ATG4B exhibit a severe but incomplete defect in LC3/GABARAP processing and autophagy. By further genetic depletion of ATG4 isoforms using CRISPR-Cas9 and siRNA we uncover that ATG4A, ATG4C and ATGD all contribute to residual priming activity, which is sufficient to enable lipidation of endogenous GABARAPL1 on autophagic structures. We also demonstrate that expressing high levels of pre-primed LC3B in ATG4-deficient cells can rescue a defect in autophagic degradation of the cargo receptor SQSTM1/p62, suggesting that delipidation by human ATG4 is not essential for autophagosome formation and fusion with lysosomes. Overall, our study provides a comprehensive characterization of ATG4 isoform function during autophagy in human cells

Searching for galactic axions through magnetized media: QUAX status report

The current status of the QUAX R\&D program is presented. QUAX is a feasibility study for a detection of axion as dark matter based on the coupling to the electrons. The relevant signal is a magnetization change of a magnetic material placed inside a resonant microwave cavity and polarized with a static magnetic field.Comment: Contributed to the 13th Patras Workshop on Axions, WIMPs and WISPs, Thessaloniki, May 15 to 19, 201

ATLAS Infrastructure

This document describes the civil engineering and infrastructure work done on the surface and underground for the ATLAS experiment at point 1 of the LHC ring

First Cool-down and Test at 4.5 K of the ATLAS Superconducting Barrel Toroid Assembled in the LHC Experimental Cavern

The large ATLAS superconducting magnets system consists of the Barrel, two End-Caps Toroids and the Central Solenoid. The eight separate coils making the Barrel Toroid (BT) have been individually tested with success in a dedicated surface test facility in 2004 and 2005 and afterwards assembled in the underground cavern of the ATLAS experiment. In order to fulfil all the cryogenic scenarios foreseen for these magnets with a cold mass of 370 tons, two separate helium refrigerators and a complex helium distribution system have been used. This paper describes the results of the first cool-down, steady-state operation at 4.5 K and quench recovery of the BT in its final configuration

The J-elongated conformation of β2-glycoprotein I predominates in solution: implications for our understanding of antiphospholipid syndrome

β2-Glycoprotein I (β2GPI) is an abundant plasma protein displaying phospholipid-binding properties. Because it binds phospholipids, it is a target of antiphospholipid antibodies (aPLs) in antiphospholipid syndrome (APS), a life-threatening autoimmune thrombotic disease. Indeed, aPLs prefer membrane-bound β2GPI to that in solution. β2GPI exists in two almost equally populated redox states: oxidized, in which all the disulfide bonds are formed, and reduced, in which one or more disulfide bonds are broken. Furthermore, β2GPI can adopt multiple conformations (i.e. J-elongated, S-twisted, and O-circular). While strong evidence indicates that the J-form is the structure bound to aPLs, which conformation exists and predominates in solution remains controversial, and so is the conformational pathway leading to the bound state. Here, we report that human recombinant β2GPI purified under native conditions is oxidized. Moreover, under physiological pH and salt concentrations, this oxidized form adopts a J-elongated, flexible conformation, not circular or twisted, in which the N-terminal domain I (DI) and the C-terminal domain V (DV) are exposed to the solvent. Consistent with this model, binding kinetics and mutagenesis experiments revealed that in solution the J-form interacts with negatively charged liposomes and with MBB2, a monoclonal anti-DI antibody that recapitulates most of the features of pathogenic aPLs. We conclude that the preferential binding of aPLs to phospholipid-bound β2GPI arises from the ability of its preexisting J-form to accumulate on the membranes, thereby offering an ideal environment for aPL binding. We propose that targeting the J-form of β2GPI provides a strategy to block pathogenic aPLs in APS