Inhibition of the EGF receptor by binding of MIG6 to an activating kinase domain interface.

Members of the epidermal growth factor receptor family (EGFR/ERBB1, ERBB2/HER2, ERBB3/HER3 and ERBB4/HER4) are key targets for inhibition in cancer therapy. Critical for activation is the formation of an asymmetric dimer by the intracellular kinase domains, in which the carboxy-terminal lobe (C lobe) of one kinase domain induces an active conformation in the other. The cytoplasmic protein MIG6 (mitogen-induced gene 6; also known as ERRFI1) interacts with and inhibits the kinase domains of EGFR and ERBB2 (refs 3-5). Crystal structures of complexes between the EGFR kinase domain and a fragment of MIG6 show that a approximately 25-residue epitope (segment 1) from MIG6 binds to the distal surface of the C lobe of the kinase domain. Biochemical and cell-based analyses confirm that this interaction contributes to EGFR inhibition by blocking the formation of the activating dimer interface. A longer MIG6 peptide that is extended C terminal to segment 1 has increased potency as an inhibitor of the activated EGFR kinase domain, while retaining a critical dependence on segment 1. We show that signalling by EGFR molecules that contain constitutively active kinase domains still requires formation of the asymmetric dimer, underscoring the importance of dimer interface blockage in MIG6-mediated inhibition

Morphological Characteristics and Allometric Relationships of Shoot in Two Undergrowth Plants: Polygonatum odoratum and Polygonatum multiflorum

The main purpose of this investigation was to describe the spatial arrangement of shoot tissues, as seen in transverse section, and allometric relationships in two contrasting species of Polygonatum i.e., Polygonatum odoratum which commonly grows in mixed pine-oak forest with shoots rectangular in shape, and Polygonatum multiflorum found in oak-hornbeam forest with cylindrical shoots. The mass and length of the aerial shoots of each individual plant were measured. The shoot regions of each plant were then categorized as basal (b), central (c) or apical (a). Transverse sections of these shoot regions were subsequently cut, and the following parameters were measured: (1) Diameter of shoots, (2) thickness of the outer and inner zones of parenchyma and (3) thickness of the sclerenchyma zone. Additional allometric relationships between the various measurements were computed and determined as Pearson’s correlation coefficients (r). Both species of Polygonatum differed significantly with respect to the length, diameter and thickness of the outer zone of parenchyma. Shoots of P. multiflorum were taller but narrower than those of P. odoratum, which had a significantly wider zone of outer parenchyma. Allometric relationships were stronger for P. multiflorum, and for both species, they were generally stronger in the basal part of the shoot. We conclude that in P. multiflorum, the strong correlation between the diameter and length of the shoot seems to be important to the growth in shaded environments

Functional role of PGAM5 multimeric assemblies and their polymerization into filaments.

PGAM5 is a mitochondrial protein phosphatase whose genetic ablation in mice results in mitochondria-related disorders, including neurodegeneration. Functions of PGAM5 include regulation of mitophagy, cell death, metabolism and aging. However, mechanisms regulating PGAM5 activation and signaling are poorly understood. Using electron cryo-microscopy, we show that PGAM5 forms dodecamers in solution. We also present a crystal structure of PGAM5 that reveals the determinants of dodecamer formation. Furthermore, we observe PGAM5 dodecamer assembly into filaments both in vitro and in cells. We find that PGAM5 oligomerization into a dodecamer is not only essential for catalytic activation, but this form also plays a structural role on mitochondrial membranes, which is independent of phosphatase activity. Together, these findings suggest that modulation of the oligomerization of PGAM5 may be a regulatory switch of potential therapeutic interest

A warped m=2 water maser disc in V778 Cyg?

The silicate carbon star V778 Cyg is a source of 22 GHz water maser emission which was recently resolved by MERLIN. Observations revealed an elongated S-like structure along which the velocities of the maser features show a linear dependence on the impact parameter. This is consistent with a doubly-warped m=2 disc observed edge-on. Water masers and silicate dust emission (detected by IRAS and ISO) have a common origin in O-rich material and are likely to be co-located in the disc. We propose a detailed self-consistent model of a masing gas-dust disc around a companion to the carbon star in a binary system, which allows us to estimate the companion mass of 1.7 +- 0.1 M_sun, the disc radius of 40 +-3 AU and the distance between companions of about 80 AU. Using a dust-gas coupling model for water masing, we calculate the maser power self-consistently, accounting for both the gas and the dust energy balances. Comparing the simulation results with the observational data, we deduce the main physical parameters of the masing disc, such as the gas and dust temperatures and their densities. We also present an analysis of the stability of the disc.Comment: 7 pages, 5 figures. This paper is accepted for publication in MNRA

hSpry2 Is Targeted to the Ubiquitin-Dependent Proteasome Pathway by c-Cbl

AbstractSprouty was originally identified in a genetic screen in Drosophila as an antagonist of fibroblast (FGF) and epidermal growth factor (EGF) signaling [1, 2]. Subsequently, four vertebrate homologs were discovered; among these, the human homolog Sprouty 2 (hSpry2) contains the highest degree of sequence homology to the Drosophila protein [3, 4]. It has been shown that hSpry2 interacts directly with c-Cbl, an E3-ubiquitin ligase, which promotes the downregulation of receptor tyrosine kinases (RTKs) [5]. In this study, we have investigated the functional consequences of the association between hSpry2 and c-Cbl. We have found that hSpry2 is ubiquitinated by c-Cbl in an EGF-dependent manner. EGF stimulation induces the tyrosine phosphorylation of hSpry2, which in turn enhances the interaction of hSpry2 with c-Cbl. The c-Cbl-mediated ubiquitination of hSpry2 targets the protein for degradation by the 26S proteasome. An enhanced proteolytic degradation of hSpry2 is also observed in response to FGF stimulation. The FGF-induced degradation of hSpry2 limits the duration of the inhibitory effect of hSpry2 on extracellular signal-regulated kinase (ERK) activation and enables the cells to recover their sensitivity to FGF stimulation. Our results indicate that the interaction of hSpry2 with c-Cbl might serve as a mechanism for the downregulation of hSpry2 during receptor tyrosine kinase signaling

CFD numerical modelling of a PV–TEG hybrid system cooled by air heat sink coupled with a single-phase inverter

This study presents full transient, three-dimensional numerical models of a PV–TEG hybrid module coupled with single-phase inverter by co-simulation. The influence of factors, such as wind speed, solar radiation intensity, or ambient temperature on the PV–TEG system, was also examined. The numerical model was implemented using Ansys software which accounted the phenomena of Thomson, Seebeck, and Joule’s heat place on the TEG system. Furthermore, its impact on total electrical efficiency was studied. The heat transfer surface of the passive heat sink and forced air circulation positively affected the total heat transfer, and therefore helped to maintain the electrical efficiency at a higher level. Simulation of the single-phase inverter with a PV–TEG system allows the determination of the power characteristics of the system in real time. The results of the study presented may provide a basis for performance optimization of a practical PV–TEG-inverter hybrid system co-design

Analysis of the role of the C-terminal tail in the regulation of the epidermal growth factor receptor

The ∼230-residue C-terminal tail of the epidermal growth factor receptor (EGFR) is phosphorylated upon activation. We examined whether this phosphorylation is affected by deletions within the tail and whether the two tails in the asymmetric active EGFR dimer are phosphorylated differently. We monitored autophosphorylation in cells using flow cytometry and found that the first ∼80 residues of the tail are inhibitory, as demonstrated previously. The entire ∼80-residue span is important for autoinhibition and needs to be released from both kinases that form the dimer. These results are interpreted in terms of crystal structures of the inactive kinase domain, including two new ones presented here. Deletions in the remaining portion of the tail do not affect autophosphorylation, except for a six-residue segment spanning Tyr 1086 that is critical for activation loop phosphorylation. Phosphorylation of the two tails in the dimer is asymmetric, with the activator tail being phosphorylated somewhat more strongly. Unexpectedly, we found that reconstitution of the transmembrane and cytoplasmic domains of EGFR in vesicles leads to a peculiar phenomenon in which kinase domains appear to be trapped between stacks of lipid bilayers. This artifactual trapping of kinases between membranes enhances an intrinsic functional asymmetry in the two tails in a dimer

Deletion of Mcpip1 in Mcpip1fl/flAlbCreMcpip1^{fl/fl}Alb^{Cre} mice recapitulates the phenotype of human primary biliary cholangitis

Primary biliary cholangitis (PBC) is an autoimmune disease characterized by progressive destruction of the intrahepatic bile ducts. The immunopathology of PBC involves excessive inflammation; therefore, negative regulators of inflammatory response, such as Monocyte Chemoattractant Protein-1-Induced Protein-1 (MCPIP1) may play important roles in the development of PBC. The aim of this work was to verify whether Mcpip1 expression protects against development of PBC. Genetic deletion of Zc3h12a was used to characterize the role of Mcpip1 in the pathogenesis of PBC in 6–52-week-old mice. We found that Mcpip1 deficiency in the liver (Mcpip1fl/flAlbCre) recapitulates most of the features of human PBC, in contrast to mice with Mcpip1 deficiency in myeloid cells (Mcpip1fl/flLysMCre mice), which present with robust myeloid cell-driven systemic inflammation. In Mcpip1fl/flAlbCre livers, intrahepatic bile ducts displayed proliferative changes with inflammatory infiltration, bile duct destruction, and fibrosis leading to cholestasis. In plasma, increased concentrations of IgG, IgM, and AMA autoantibodies (anti-PDC-E2) were detected. Interestingly, the phenotype of Mcpip1fl/flAlbCre mice was robust in 6-week-old, but milder in 12–24-week-old mice. Hepatic transcriptome analysis of 6-week-old and 24-week-old Mcpip1fl/flAlbCre mice showed 812 and 8 differentially expressed genes, respectively, compared with age-matched control mice, and revealed a distinct set of genes compared to those previously associated with development of PBC. In conclusion, Mcpip1fl/flAlbCre mice display early postnatal phenotype that recapitulates most of the features of human PBC

A Novel Monoallelic Nonsense Mutation in the NFKB2 Gene Does Not Cause a Clinical Manifestation

NF-κB signaling, acting through NFKB1 dependent canonical and NFKB2 dependent non-canonical pathways plays a critical role in inflammatory and immune responses. Recent studies have associated mutations in these two genes with a common variable immunodeficiency (CVID). While evaluating a female patient seeking a diagnosis explaining her recurrent infections, we found a novel heterozygous c.1831C > T (p.Arg611∗) nonsense mutation in the NFKB2 gene which introduces a Stop codon in the ankyrin repeat domain of p100. Whole exome sequencing (WES) analysis, followed by Sanger sequencing, identified this previously unknown mutation in two other family members. Penetrance of the c.1831C > T variant was assessed by flow-cytometry and protein expression in peripheral blood mononuclear cells (PBMC); whereas, activation of the NF-κB2 signaling pathway was examined through immunoblotting and real-time PCR. Heterozygous c.1831C > T variant led to the expansion of lymphocyte B subpopulations with concomitant reduction of plasmablasts, low IgG levels, and accumulation of p52 in PBMC. On the other hand, tested subjects had normal levels of IgM, IgA, IgE and no impairment in lymphocytes proliferation. Although evaluated patients did not fulfill all clinical features of CVID, their health should be monitored in the future for possible late manifestation of the disease. In conclusion, we showed that NFKB2 haplodeficiency caused by c.1831C > T nonsense mutation is asymptomatic, possibly due to the compensatory mechanisms and allele redundancy