Functional interaction between the ZO-1-interacting transcription factor ZONAB/DbpA and the RNA processing factor symplekin

Epithelial tight junctions participate in the regulation of gene expression by controlling the activity of transcription factors that can interact with junctional components. One such protein is the Y-box transcription factor ZONAB/DbpA that binds to ZO-1, a component of the junctional plaque. Symplekin, another nuclear protein that can associate with tight junctions, functions in the regulation of polyadenylation and thereby promotes gene expression. Here, we addressed the question of whether these two proteins interact and whether this is of functional relevance. We demonstrate that ZONAB/DbpA and symplekin form a complex in kidney and intestinal epithelial cells that can be immunoprecipitated and that exists in the nucleus. The interaction between ZONAB/DbpA and symplekin can be reconstituted with recombinant proteins. In reporter gene assays in which ZONAB/DbpA functions as a repressor, symplekin functionally interacts with ZONAB/DbpA, indicating that symplekin can also promote transcriptional repression. RNAi experiments indicate that symplekin depletion reduces the nuclear accumulation and the transcriptional activity of ZONAB/DbpA in colon adenocarcinoma cells, resulting in inhibition of proliferation and reduced expression of the ZONAB/DbpA-target gene cyclin D1. Our data thus indicate that symplekin and ZONAB/DbpA cooperate in the regulation of transcription, and that they promote epithelial proliferation and cyclin D1 expression

Finite element analysis of fire resistant reinforcement on end-plate steel connections

In this paper the effect of fire resistant coatings on the mechanical behaviour of steel joints is studied using the finite element method. The proposed finite element model is an extension of a previous one developed for the study of the same connection in elevated temperatures, without fire reinforcement. In particular, the construction used consists of an end – plate steel connection which is covered with panels of lightweight concrete and gypsum board. The behaviour of those two fire resistant materials has been simulated in elevating mechanical and thermal conditions separately and simultaneously. Through this process it is examined the strength of the materials and of the overall construction. Specifically, the action of fire on the strength of the structure may result in an early collapse. In addition, the behaviour of the structure in the connection area and the opening of the interface is investigated

Annexin A1 Tethers Membrane Contact Sites that Mediate ER to Endosome Cholesterol Transport

Membrane contact sites between the ER and multivesicular endosomes/bodies (MVBs) play important roles in endosome positioning and fission and in neurite outgrowth. ER-MVB contacts additionally function in epidermal growth factor receptor (EGFR) tyrosine kinase downregulation by providing sites where the ER-localized phosphatase, PTP1B, interacts with endocytosed EGFR before the receptor is sorted onto intraluminal vesicles (ILVs). Here we show that these contacts are tethered by annexin A1 and its Ca2+-dependent ligand, S100A11, and form a subpopulation of differentially regulated contact sites between the ER and endocytic organelles. Annexin A1-regulated contacts function in the transfer of ER-derived cholesterol to the MVB when low-density lipoprotein-cholesterol in endosomes is low. This sterol traffic depends on interaction between ER-localized VAP and endosomal oxysterol-binding protein ORP1L, and is required for the formation of ILVs within the MVB and thus for the spatial regulation of EGFR signaling

HIGH-RESOLUTION GEOMORPHOLOGICAL MAPPING OF THE SHALLOW CONTINENTAL SHELF WEST OF THE KAVALA BAY, NORTH AEGEAN

Σημαντικές γεωμορφολογικές δομές του ρηχού τμήματος της υφαλοκρηπίδας δυτικά του Κόλπου της Καβάλας χαρτογραφήθηκαν χρησιμοποιώντας τα δεδομένα από μια υδρογραφική αποτύπωση (τον Ιούνιο 2014) 320 ναυτικών μιλίων, η οποία περιελάμβανε υψηλής διακριτικότητας πολυδεσμική βαθυμετρική καταγραφή και διασκόπηση πυθμένα με σεισμική ανάκλαση. Αναγνωρίστηκε ένα σύστημα ρηγμάτων αποτελούμενο από ένα σετ δυο κυρίων κανονικών ρηγμάτων (καταγεγραμμένο μήκος και μετρημένο κατακόρυφο άλμα αυτών: 12 χλμ, 5 χλμ και > 40 μ, 25 μ, αντίστοιχα,) με έντονη επιφανειακή εκδήλωση στο θαλάσσιο πυθμένα, καθώς και τρία δευτερεύοντα ρήγματα νότια των κύριων ρηγμάτων, τα οποία φανερώνουν συνιζηματογενή τεκτονισμό. Η εντυπωσιακή διαφορά στις υφές των ιζημάτων που καλύπτουν αφενός το υποκείμενο ρηξιτέμαχος του βορειότερου κυρίου ρήγματος και αφετέρου την οροφή του νοτιότερου κυρίου ρήγματος δείχνει τη σημαντική επίδραση του τεκτονισμού στις ιζηματολογκές διεργασίες της περιοχής μελέτης. Όσον αφορά τις υπάρχουσες γεωμορφές, οι περισσότερο ενδιαφέρουσες είναι εκείνες των αμμωδών θινών στο βορειοανατολικό τμήμα της περιοχής μελέτης, ευρισκόμενες σε βάθη από 25 μ μέχρι τουλάχιστον 65 μ. Οι μεγάλες διαστάσεις τους καθώς και ο προσανατολισμός τους ως προς την ακτογραμμή υποδηλώνουν ως μηχανισμό σχηματισμού τους την δράση ισχυρών πυθμιαίων ρευμάτωνProminent geomorphological features of the shallow continental shelf west of the Kavala Bay (Loutra Eleftheron-Nea Peramos) were mapped using the data from a hydrographic survey (June 2014) of 320 nautical miles during which high resolution multibeam bathymetry and seismic-reflection subbottom profiling were carried out simultaneously. A fault zone comprised by a set of two primary sigmoidal gravity faults (recorded lengths and measured offsets: 12 km, 5 km and > 40 m, 25 m, respectively), with distinct expression on the seabed, and three other secondary gravity faults situated southern of the major faults, revealing synsedimentary tectonics, was identified. The striking difference between the texture of the footwall block sediments of the northern major fault and the texture of the sediments occupying the deep hanging wall block of the southern major fault emphasizes the impact of local tectonics on the sedimentary evolution of the study area. Concerning the observed bedforms, the most interesting were the sand dunes occurring at depths from 25 m to 65 m at least and occupying the northeast part of the study area. Their large dimensions and orientation in relation to the coastline position imply as a mechanism for their formation intense bottom-current activity

TGF-b2 induction regulates invasiveness of theileria-transformed leukocytes and disease susceptibility

Theileria parasites invade and transform bovine leukocytes causing either East Coast fever (T. parva), or tropical theileriosis (T. annulata). Susceptible animals usually die within weeks of infection, but indigenous infected cattle show markedly reduced pathology, suggesting that host genetic factors may cause disease susceptibility. Attenuated live vaccines are widely used to control tropical theileriosis and attenuation is associated with reduced invasiveness of infected macrophages in vitro. Disease pathogenesis is therefore linked to aggressive invasiveness, rather than uncontrolled proliferation of Theileria-infected leukocytes. We show that the invasive potential of Theileria-transformed leukocytes involves TGF-b signalling. Attenuated live vaccine lines express reduced TGF-b2 and their invasiveness can be rescued with exogenous TGF-b. Importantly, infected macrophages from disease susceptible Holstein-Friesian (HF) cows express more TGF-b2 and traverse Matrigel with great efficiency compared to those from disease-resistant Sahiwal cattle. Thus, TGF-b2 levels correlate with disease susceptibility. Using fluorescence and time-lapse video microscopy we show that Theileria-infected, disease-susceptible HF macrophages exhibit increased actin dynamics in their lamellipodia and podosomal adhesion structures and develop more membrane blebs. TGF-b2-associated invasiveness in HF macrophages has a transcription-independent element that relies on cytoskeleton remodelling via activation of Rho kinase (ROCK). We propose that a TGF-b autocrine loop confers an amoeboid-like motility on Theileria-infected leukocytes, which combines with MMP-dependent motility to drive invasiveness and virulence

Bves Modulates Tight Junction Associated Signaling

Blood vessel epicardial substance (Bves) is a transmembrane adhesion protein that regulates tight junction (TJ) formation in a variety of epithelia. The role of TJs within epithelium extends beyond the mechanical properties. They have been shown to play a direct role in regulation of RhoA and ZONAB/DbpA, a y-box transcription factor. We hypothesize that Bves can modulate RhoA activation and ZONAB/DbpA activity through its regulatory effect on TJ formation. Immortalized human corneal epithelial (HCE) cells were stably transfected with Flag-tagged full length chicken Bves (w-Bves) or C-terminus truncated Bves (t-Bves). We found that stably transfected w-Bves and t-Bves were interacting with endogenous human Bves. However, interaction with t-Bves appeared to disrupt cell membrane localization of endogenous Bves and interaction with ZO-1. w-Bves cells exhibited increased TJ function reflected by increased trans-epithelial electrical resistance, while t-Bves cells lost TJ protein immunolocalization at cell-cell contacts and exhibited decreased trans-epithelial electrical resistance. In parental HCE and w-Bves cells ZONAB/DbpA and GEF-H1 were seen at cell borders in the same pattern as ZO-1. However, expression of t-Bves led to decreased membrane localization of both ZONAB/DbpA and GEF-H1. t-Bves cells had increased RhoA activity, as indicated by a significant 30% increase in FRET activity compared to parental HCE cells. ZONAB/DbpA transcriptional activity, assessed using a luciferase reporter probe, was increased in t-Bves cells. These studies demonstrate that Bves expression and localization can regulate RhoA and ZONAB/DbpA activity

The Tight Junction Associated Signalling Proteins ZO-1 and ZONAB Regulate Retinal Pigment Epithelium Homeostasis in Mice

Cell-cell adhesion regulates the development and function of epithelia by providing mechanical support and by guiding cell proliferation and differentiation. The tight junction (TJ) protein zonula occludens (ZO)-1 regulates cell proliferation and gene expression by inhibiting the activity of the Y-box transcription factor ZONAB in cultured epithelial cells. We investigated the role of this TJ-associated signalling pathway in the retinal pigment epithelium (RPE) in vivo by lentivirally-mediated overexpression of ZONAB, and knockdown of its cellular inhibitor ZO-1. Both overexpression of ZONAB or knockdown of ZO-1 resulted in increased RPE proliferation, and induced ultrastructural changes of an epithelial-mesenchymal transition (EMT)-like phenotype. Electron microscopy analysis revealed that transduced RPE monolayers were disorganised with increased pyknosis and monolayer breaks, correlating with increased expression of several EMT markers. Moreover, fluorescein angiography analysis demonstrated that the increased proliferation and EMT-like phenotype induced by overexpression of ZONAB or downregulation of ZO-1 resulted in RPE dysfunction. These findings demonstrate that ZO-1 and ZONAB are critical for differentiation and homeostasis of the RPE monolayer and may be involved in RPE disorders such as proliferative vitroretinopathy and atrophic age-related macular degeneration

Claudins in lung diseases

Tight junctions are the most apically localized part of the epithelial junctional complex. They regulate the permeability and polarity of cell layers and create compartments in cell membranes. Claudins are structural molecules of tight junctions. There are 27 claudins known, and expression of different claudins is responsible for changes in the electrolyte and solute permeability in cells layers. Studies have shown that claudins and tight junctions also protect multicellular organisms from infections and that some infectious agents may use claudins as targets to invade and weaken the host's defense. In neoplastic diseases, claudin expression may be up- or downregulated. Since their expression is associated with specific tumor types or with specific locations of tumors to a certain degree, they can, in a restricted sense, also be used as tumor markers. However, the regulation of claudin expression is complex involving growth factors and integrins, protein kinases, proto-oncogens and transcription factors. In this review, the significance of claudins is discussed in lung disease and development

Organization of multiprotein complexes at cell–cell junctions

The formation of stable cell–cell contacts is required for the generation of barrier-forming sheets of epithelial and endothelial cells. During various physiological processes like tissue development, wound healing or tumorigenesis, cellular junctions are reorganized to allow the release or the incorporation of individual cells. Cell–cell contact formation is regulated by multiprotein complexes which are localized at specific structures along the lateral cell junctions like the tight junctions and adherens junctions and which are targeted to these site through their association with cell adhesion molecules. Recent evidence indicates that several major protein complexes exist which have distinct functions during junction formation. However, this evidence also indicates that their composition is dynamic and subject to changes depending on the state of junction maturation. Thus, cell–cell contact formation and integrity is regulated by a complex network of protein complexes. Imbalancing this network by oncogenic proteins or pathogens results in barrier breakdown and eventually in cancer. Here, I will review the molecular organization of the major multiprotein complexes at junctions of epithelial cells and discuss their function in cell–cell contact formation and maintenance