The deubiquitinase Leon/USP5 interacts with Atg1/ULK1 and antagonizes autophagy.

Accumulating evidence has shown that the quality of proteins must be tightly monitored and controlled to maintain cellular proteostasis. Misfolded proteins and protein aggregates are targeted for degradation through the ubiquitin proteasome (UPS) and autophagy-lysosome systems. The ubiquitination and deubiquitinating enzymes (DUBs) have been reported to play pivotal roles in the regulation of the UPS system. However, the function of DUBs in the regulation of autophagy remain to be elucidated. In this study, we found that knockdown of Leon/USP5 caused a marked increase in the formation of autophagosomes and autophagic flux under well-fed conditions. Genetic analysis revealed that overexpression of Leon suppressed Atg1-induced cell death in Drosophila. Immunoblotting assays further showed a strong interaction between Leon/USP5 and the autophagy initiating kinase Atg1/ULK1. Depletion of Leon/USP5 led to increased levels of Atg1/ULK1. Our findings indicate that Leon/USP5 is an autophagic DUB that interacts with Atg1/ULK1, negatively regulating the autophagic process.

Control of Large-Area Orderliness of a 2D Supramolecular Chiral Microstructure by a 1D Interference Field.

Self-organized periodic micro/nanostructures caused by stimulus-responsive structural deformation often occur in anisotropic self-assembled supramolecular systems (e.g., liquid crystal systems). However, the long-range orderliness of these structures is often beyond control. In this article, we first demonstrate that a large-area disordered two-dimensional (2D) microgrid chiral structure appears in the cholesteric liquid crystal (CLC) reactive mixture because of the photopolymerization-induced Helfrich deformation effect under exposure to the single UV-laser beam. The result is attributed to the impact of an internal longitudinal strain, which is caused by the pitch contraction of the CLC-monomer region through the continuing compression of the thickening CLC polymer layer adhered on the illuminated substrate of the sample during photopolymerization. The experimental results further show that a one-dimensional (1D) UV-laser interference field can be used to effectively control the postformed 2D microgrid structure to arrange in an orderly manner throughout the large exposed area (an order of centimeter). The optimum ability for controlling the orderliness of the microgrid structure can be achieved if the spacing width of the interference field approximates the periodicity of the postformed 2D microgrids. Several factors, such as the pitch of the CLC mixture and the included angle and intensity of the two interfering laser beams, which influence the orderliness and properties of the 2D microgrid structure, are explored in this study. The result of this research opens a new page to improve the applicability of the Helfrich deformation phenomenon and further provides a reference platform for manipulating, modifying, and even tailoring periodic micro/nanostructures in self-organized supramolecular soft-matter systems for application in advanced optics/photonics.

USP5/Leon deubiquitinase confines postsynaptic growth by maintaining ubiquitin homeostasis through Ubiquilin.

Synapse formation and growth are tightly controlled processes. How synaptic growth is terminated after reaching proper size remains unclear. Here, we show that Leon, the Drosophila USP5 deubiquitinase, controls postsynaptic growth. In leon mutants, postsynaptic specializations of neuromuscular junctions are dramatically expanded, including the subsynaptic reticulum, the postsynaptic density, and the glutamate receptor cluster. Expansion of these postsynaptic features is caused by a disruption of ubiquitin homeostasis with accumulation of free ubiquitin chains and ubiquitinated substrates in the leon mutant. Accumulation of Ubiquilin (Ubqn), the ubiquitin receptor whose human homolog ubiquilin 2 is associated with familial amyotrophic lateral sclerosis, also contributes to defects in postsynaptic growth and ubiquitin homeostasis. Importantly, accumulations of postsynaptic proteins cause different aspects of postsynaptic overgrowth in leon mutants. Thus, the deubiquitinase Leon maintains ubiquitin homeostasis and proper Ubqn levels, preventing postsynaptic proteins from accumulation to confine postsynaptic growth.

Dbo/Henji Modulates Synaptic dPAK to Gate Glutamate Receptor Abundance and Postsynaptic Response.

In response to environmental and physiological changes, the synapse manifests plasticity while simultaneously maintains homeostasis. Here, we analyzed mutant synapses of henji, also known as dbo, at the Drosophila neuromuscular junction (NMJ). In henji mutants, NMJ growth is defective with appearance of satellite boutons. Transmission electron microscopy analysis indicates that the synaptic membrane region is expanded. The postsynaptic density (PSD) houses glutamate receptors GluRIIA and GluRIIB, which have distinct transmission properties. In henji mutants, GluRIIA abundance is upregulated but that of GluRIIB is not. Electrophysiological results also support a GluR compositional shift towards a higher IIA/IIB ratio at henji NMJs. Strikingly, dPAK, a positive regulator for GluRIIA synaptic localization, accumulates at the henji PSD. Reducing the dpak gene dosage suppresses satellite boutons and GluRIIA accumulation at henji NMJs. In addition, dPAK associated with Henji through the Kelch repeats which is the domain essential for Henji localization and function at postsynapses. We propose that Henji acts at postsynapses to restrict both presynaptic bouton growth and postsynaptic GluRIIA abundance by modulating dPAK.

The deubiquitinase Leon/USP5 regulates ubiquitin homeostasis during Drosophila development.

Ubiquitination and the reverse process deubiquitination regulate protein stability and function during animal development. The Drosophila USP5 homolog Leon functions as other family members of unconventional deubiquitinases, disassembling free, substrate-unconjugated polyubiquitin chains to replenish the pool of mono-ubiquitin, and maintaining cellular ubiquitin homeostasis. However, the significance of Leon/USP5 in animal development is still unexplored. In this study, we generated leon mutants to show that Leon is essential for animal viability and tissue integrity during development. Both free and substrate-conjugated polyubiquitin chains accumulate in leon mutants, suggesting that abnormal ubiquitin homeostasis caused tissue disorder and lethality in leon mutants. Further analysis of protein expression profiles in leon mutants shows that the levels of all proteasomal subunits were elevated. Also, proteasomal enzymatic activities were elevated in leon mutants. However, proteasomal degradation of ubiquitinated substrates was impaired. Thus, aberrant ubiquitin homeostasis in leon mutants disrupts normal proteasomal degradation, which is compensated by elevating the levels of proteasomal subunits and activities. Ultimately, the failure to fully compensate the dysfunctional proteasome in leon mutants leads to animal lethality and tissue disorder.

Emphysematous pyelonephritis: patient characteristics and management approach.

INTRODUCTION: Emphysematous pyelonephritis (EPN) is an acute, severe, necrotizing infection of the renal parenchyma and perirenal tissue that requires immediate treatment. However, the ideal approach to its management remains controversial. We conducted this study to determine the appropriate treatment modalities. MATERIALS AND METHODS: A retrospective review of EPN cases revealed 10 consecutive cases from July 2003 to June 2012. Clinical and demographic data were collected from each patient. RESULTS: All patients had diabetes mellitus, 5 presented with urinary tract obstruction by urolithiasis. Seven patients had type I disease and 3 had type II disease. Six of the type I patients underwent emergent nephrectomy and 1 of these died, the remaining patient refused surgical intervention and died after receiving medical management only. The type II patients underwent percutaneous drainage, and 2 of them subsequently underwent elective nephrectomy; all 3 survived. CONCLUSION: Our results suggest that emergency nephrectomy may be considered the initial management for type I EPN, while percutaneous drainage may be an effective initial treatment option for type II EPN.

Nak regulates localization of clathrin sites in higher-order dendrites to promote local dendrite growth.

VIDEO ABSTRACT: During development, dendrites arborize in a field several hundred folds of their soma size, a process regulated by intrinsic transcription program and cell adhesion molecule (CAM)-mediated interaction. However, underlying cellular machineries that govern distal higher-order dendrite extension remain largely unknown. Here, we show that Nak, a clathrin adaptor-associated kinase, promotes higher-order dendrite growth through endocytosis. In nak mutants, both the number and length of higher-order dendrites are reduced, which are phenocopied by disruptions of clathrin-mediated endocytosis. Nak interacts genetically with components of the endocytic pathway, colocalizes with clathrin puncta, and is required for dendritic localization of clathrin puncta. More importantly, these Nak-containing clathrin structures preferentially localize to branching points and dendritic tips that are undergoing active growth. We present evidence that the Drosophila L1-CAM homolog Neuroglian is a relevant cargo of Nak-dependent internalization, suggesting that localized clathrin-mediated endocytosis of CAMs facilitates the extension of nearby higher-order dendrites.

Suppression of Hedgehog signaling by Cul3 ligases in proliferation control of retinal precursors.

Cullin-RING ubiquitin ligases ubiquitinate protein substrates and control their levels through degradation. Here we show that cullin3 (Cul3) suppresses Hedgehog (Hh) signaling through downregulating the level of the signaling pathway effector cubitus interruptus (Ci). High-level Hh signaling promotes Cul3-dependent Ci degradation, leading to the downregulation of Hh signaling. This process is manifested in controlling cell proliferation during Drosophila retinal development. In Cul3 mutants, the population of interommatidial cells is increased, which can be mimicked by overexpression of Ci and suppressed by depleting endogenous Ci. Hh also regulates the population of interommatidial cells in the pupal stage. Alterations in the interommatidial cell population correlate with alterations in precursor proliferation in the second mitotic wave of larval eye discs. Taken together, these results suggest that Cul3 downregulates Ci levels to modulate Hh signaling activity, thus ensuring proper cell proliferation during retinal development.

Testicular metastasis from hepatocellular carcinoma.

We document a case of testicular metastasis from hepatocellular carcinoma. The patient suffered from bilateral testicular painful swelling for 6 months. Scrotal ultrasonography showed bilateral testicular tumors and the whole abdominal computed tomography revealed a huge tumor in the left lobe of the liver. Bilateral orchiectomy and postoperative ultrasound-guided liver biopsy were done. Pathological examination revealed metastatic hepatocellular carcinoma. Hepatocellular carcinoma with testicular metastases is a very rare disease.