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

Pitx2 cholinergic interneurons are the source of C bouton synapses on brainstem motor neurons

IR and LZ were funded by the European Union, Seventh Framework Programme (FP7/2007–2013), by the European Union and Greek National Funds through the operational program “Education and Lifelong Learning” of the National Strategic Reference Framework, funding program: ARISTEIA II, and by Fondation Santé.Cholinergic neuromodulation has been described throughout the brain and has been implicated in various functions including attention, food intake and response to stress. Cholinergic modulation is also thought to be important for regulating motor systems, as revealed by studies of large cholinergic synapses on spinal motor neurons, called C boutons, which seem to control motor neuron excitability in a task-dependent manner. C boutons on spinal motor neurons stem from spinal interneurons that express the transcription factor Pitx2. C boutons have also been identified on the motor neurons of specific cranial nuclei. However, the source and roles of cranial C boutons are less clear. Previous studies suggest that they originate from Pitx2+ and Pitx2− neurons, in contrast to spinal cord C boutons that originate solely from Pitx2 neurons. Here, we address this controversy using mouse genetics, and demonstrate that brainstem C boutons are Pitx2+ derived. We also identify new Pitx2 populations and map the cholinergic Pitx2 neurons of the mouse brain. Taken together, our data present important new information about the anatomical organization of cholinergic systems which impact motor systems of the brainstem. These findings will enable further analyses of the specific roles of cholinergic modulation in motor control.Publisher PDFPeer reviewe

Matrix stiffening sensitizes epithelial cells to EGF and enables the loss of contact inhibition of proliferation

Anchorage to a compliant extracellular matrix (ECM) and contact with neighboring cells impose important constraints on the proliferation of epithelial cells. How anchorage and contact dependence are inter-related and how cells weigh these adhesive cues alongside soluble growth factors to make a net cell cycle decision remain unclear. Here, we show that a moderate 4.5-fold stiffening of the matrix reduces the threshold amount of epidermal growth factor (EGF) needed to over-ride contact inhibition by over 100-fold. At EGF doses in the range of the dissociation constant (Kd) for ligand binding, epithelial cells on soft matrices are contact inhibited with DNA synthesis restricted to the periphery of cell clusters. By contrast, on stiff substrates, even EGF doses at sub-Kd levels over-ride contact inhibition, leading to proliferation throughout the cluster. Thus, matrix stiffening significantly sensitizes cells to EGF, enabling contact-independent spatially uniform proliferation. Contact inhibition on soft substrates requires E-cadherin, and the loss of contact inhibition upon matrix stiffening is accompanied by the disruption of cell–cell contacts, changes in the localization of the EGF receptor and ZO-1, and selective attenuation of ERK, but not Akt, signaling. We propose a quantitative framework for the epigenetic priming (via ECM stiffening) of a classical oncogenic pathway (EGF) with implications for the regulation of tissue growth during morphogenesis and cancer progression

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