The α-arrestin ARRDC3 mediates ALIX ubiquitination and G protein-coupled receptor lysosomal sorting.

The sorting of G protein-coupled receptors (GPCRs) to lysosomes is critical for proper signaling and cellular responses. We previously showed that the adaptor protein ALIX regulates lysosomal degradation of protease-activated receptor-1 (PAR1), a GPCR for thrombin, independent of ubiquitin-binding ESCRTs and receptor ubiquitination. However, the mechanisms that regulate ALIX function during PAR1 lysosomal sorting are not known. Here we show that the mammalian α-arrestin arrestin domain-containing protein-3 (ARRDC3) regulates ALIX function in GPCR sorting via ubiquitination. ARRDC3 colocalizes with ALIX and is required for PAR1 sorting at late endosomes and degradation. Depletion of ARRDC3 by small interfering RNA disrupts ALIX interaction with activated PAR1 and the CHMP4B ESCRT-III subunit, suggesting that ARRDC3 regulates ALIX activity. We found that ARRDC3 is required for ALIX ubiquitination induced by activation of PAR1. A screen of nine mammalian NEDD4-family E3 ubiquitin ligases revealed a critical role for WWP2. WWP2 interacts with ARRDC3 and not ALIX. Depletion of WWP2 inhibited ALIX ubiquitination and blocked ALIX interaction with activated PAR1 and CHMP4B. These findings demonstrate a new role for the α-arrestin ARRDC3 and the E3 ubiquitin ligase WWP2 in regulation of ALIX ubiquitination and lysosomal sorting of GPCRs

The HTLV Receptor Is a Widely Expressed Protein

AbstractThe receptor for human T-cell leukemia virus type 1 (HTLV-1) was found to be expressed on a broad range of cell lines derived from multiple species. Receptor expression was assessed using human immunodeficiency virus type 1 particles, pseudotyped with the HTLV-1 envelope glycoprotein, and expressing luciferase under the control of an SV40 enhancer and promoter. Infection by pseudotyped virus was blocked with neutralizing antibodies to HTLV-1, and infection was dependent on the presence of the cleavage and fusogenic sequences in the envelope protein precursor. Trypsin treatment of susceptible target lymphocytes reduced entry. Entry was partially resistant to ammonium chloride

Insulin secretion in health and disease: nutrients dictate the pace.

Insulin is a key hormone controlling metabolic homeostasis. Loss or dysfunction of pancreatic β-cells lead to the release of insufficient insulin to cover the organism needs, promoting diabetes development. Since dietary nutrients influence the activity of β-cells, their inadequate intake, absorption and/or utilisation can be detrimental. This review will highlight the physiological and pathological effects of nutrients on insulin secretion and discuss the underlying mechanisms. Glucose uptake and metabolism in β-cells trigger insulin secretion. This effect of glucose is potentiated by amino acids and fatty acids, as well as by entero-endocrine hormones and neuropeptides released by the digestive tract in response to nutrients. Glucose controls also basal and compensatory β-cell proliferation and, along with fatty acids, regulates insulin biosynthesis. If in the short-term nutrients promote β-cell activities, chronic exposure to nutrients can be detrimental to β-cells and causes reduced insulin transcription, increased basal secretion and impaired insulin release in response to stimulatory glucose concentrations, with a consequent increase in diabetes risk. Likewise, suboptimal early-life nutrition (e.g. parental high-fat or low-protein diet) causes altered β-cell mass and function in adulthood. The mechanisms mediating nutrient-induced β-cell dysfunction include transcriptional, post-transcriptional and translational modifications of genes involved in insulin biosynthesis and secretion, carbohydrate and lipid metabolism, cell differentiation, proliferation and survival. Altered expression of these genes is partly caused by changes in non-coding RNA transcripts induced by unbalanced nutrient uptake. A better understanding of the mechanisms leading to β-cell dysfunction will be critical to improve treatment and find a cure for diabetes

ALIX binds a YPX(3)L motif of the GPCR PAR1 and mediates ubiquitin-independent ESCRT-III/MVB sorting.

The sorting of signaling receptors to lysosomes is an essential regulatory process in mammalian cells. During degradation, receptors are modified with ubiquitin and sorted by endosomal sorting complex required for transport (ESCRT)-0, -I, -II, and -III complexes into intraluminal vesicles (ILVs) of multivesicular bodies (MVBs). However, it remains unclear whether a single universal mechanism mediates MVB sorting of all receptors. We previously showed that protease-activated receptor 1 (PAR1), a G protein-coupled receptor (GPCR) for thrombin, is internalized after activation and sorted to lysosomes independent of ubiquitination and the ubiquitin-binding ESCRT components hepatocyte growth factor-regulated tyrosine kinase substrate and Tsg101. In this paper, we report that PAR1 sorted to ILVs of MVBs through an ESCRT-III-dependent pathway independent of ubiquitination. We further demonstrate that ALIX, a charged MVB protein 4-ESCRT-III interacting protein, bound to a YPX(3)L motif of PAR1 via its central V domain to mediate lysosomal degradation. This study reveals a novel MVB/lysosomal sorting pathway for signaling receptors that bypasses the requirement for ubiquitination and ubiquitin-binding ESCRTs and may be applicable to a subset of GPCRs containing YPX(n)L motifs

Assessment of the diagnostic accuracy of bacteriological culture and the invA-gen-based PCR for the detection of Salmonella organisms from caecal content from slaughtered pigs through Bayesian approaches

The goal of this study was to determine the accuracy of a culture technique and the invA-gen-based PCR, for the detection of Salmonella spp. From caecal samples from slaughtered pigs. For this purpose a Bayesian approach was used. Two hundred and three pigs were used. Animals were grouped into 2 populations: 96 from small farms and 107 from large farms. Sensitivity was 56% (95% Credible Interval: 40-76) for culture and 91% (95% CI: 81-97) for PCR

A Tale of Two Sylamores: Understanding Relationships Among Land Use, Nutrients, and Aquatic Communities Across a Subsidy-Stress Gradient

Agricultural land use is known to degrade aquatic systems with high inputs of nutrients, sediments, and pesticides. Increased nutrients can lead to increased algal growth and thus possible hypoxic conditions in slow moving water, while increased sediment loads have been shown to obstruct light and reduce substrate stability. These conditions negatively impact primary producers, macroinvertebrates, and fish. However, small-scale changes in land use can subsidize an aquatic ecosystem instead, where an increase in nutrients allows nutrient-limited biota to flourish, and minor increases in sedimentation may help support populations of collector-filterers. The stimulation in performance caused by small disturbances is part of the subsidy-stress gradient, where increasing perturbation subsidizes an ecosystem until a certain threshold is reached, at which a decline in performance and increased variability starts to occur. The North and South Sylamore watersheds in north Arkansas provide a useful template to investigate the subsidy-stress gradient in relation to land use. North Sylamore flows through the Ozark National Forest and has a heavily forested catchment, while South Sylamore flows through mostly private land, some of which is pasture (23%). Physicochemical, macroinvertebrate, and fish data were collected from multiple sites within each watershed to determine if South Sylamore is exhibiting a response to pasture/agriculture characteristic of a subsidy-stress gradient. Sites within South Sylamore had significantly higher nitrate levels, larger macroinvertebrate populations dominated by collector-filterers, and greater abundance of algivorous fish, suggesting South Sylamore may be subsidized by the surrounding pastoral lands. However, South Sylamore also had a significantly lower proportional abundance of sensitive macroinvertebrate taxa and more unique tolerant fish taxa, suggesting South Sylamore is experiencing stress as well. Habitat quality of South Sylamore could be improved by restoration of trees within the riparian zone. Monitoring aquatic systems for subsidy-stress responses can inform restoration/management decisions and guide intervention prior to watersheds and aquatic communities becoming overly stressed