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Session XVII : Cell signaling in health and disease Poster XVII, 31 EFFECT OF ENDOTHELIN ON THE SODIUM/HYDROGEN EXCHANGER (NHE) ACTIVITY OF HUMAN MONOCYTES AND ATHEROSCLEROSIS-RELATED FUNCTIONS Koliakos G1., Befani Chr. 2, Paletas K.3, Kaloyianni M.2 1Laboratory of Biological Chemistry, Medical School, Aristotle University of Thessaloniki, 54124 2Laboratory of Animal Physiology, Department of Zoology, School of Biology, Aristotle University of Thessaloniki, 54124 3 B’ Medical Clinic, Medical School, Aristotle University of Thessaloniki, 54124 Aims: The aim of the present study is to investigate the influence of endothelin on human monocyte sodium/hydrogen exchanger (NHE-1) activity and on the atherosclerosis-related monocyte functions. NHE-1 is an integral membrane protein that exchanges one intracellular H+ ion for an extracellular Na+ ion. It plays an essential role in all cell types regulating the internal pH (pHi), cell growth, differentiation and apoptosis. Endothelin is a potent vasoconstrictive and mitogenic 21-amino-acid peptide that has been implicated in the pathogenesis of atherosclerosis. Monocytes are key contributors to the atherosclerotic process. Methods: The effect of endothelin (10pg/ml) on monocyte NHE-1 activity was studied by means of pHi (using the fluorescent indicator BCECF-AM) and 22Na uptake. Specific inhibitors were used in order to avoid interference from other sodium-exchanging pumps. In an attempt to investigate the signal transduction pathway from endothelin to NHE 1 we used PD 98059 MEK1 (50µ)) (MAPKp42/44 inhibitor), GF-109203X (10µ)) (inhibitor of all the isoforms of PKC) and Gö 6976 (500nM) (specific inhibitor of the isoform PKCs). In addition, the effect of endothelin (10pg/ml) on the ability of monocytes to bind on the basement membrane glycoprotein laminin and to migrate on trans-well culture inserts was estimated. We also measured the density of CD36 receptor using a fluorescein isothiocyanate (FITC)-linked anti CD36 monoclonal antibody. Results: Endothelin caused an increase in pHi (p
Session XVII : Cell signaling in health and disease Poster XVII, 32 Implication of TRIP6, a novel molecular partner of the MAGI-1b scaffolding molecule in invasiveness Larissa Kotelevets1, Erik Bruyneel3, Alexei Kruglov1,4, Marc Bracke3, Frans van Roy2 and Eric Chastre1 1 INSERM U683, Faculté de Médecine X. Bichat, Paris, France 2 Department for Molecular Biomedical Research, VIB-Ghent University, B-9052 Ghent 3 Laboratory of Experimental Cancerology, Ghent University Hospital, B-9000 Ghent, Belgium 4 Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino, Russia. We recently established the critical role of the PTEN/MAGI-1b signalosome in the stabilization of cell-cell contacts and the suppression of invasiveness. The PTEN tumor suppressor is recruited to E-cadherin junctional complexes through the binding to the 2nd PDZ domain of the MAGI-1b scaffolding molecule, whereas #-catenin interacts with the 5th PDZ domain of MAGI-1b (Kotelevets et al, FASEB J 19,115,2005). To identify additional effector molecules that might regulate the activity and the composition of the PTEN / MAGI- 1b complexes, we used yeast-two hybrid screening. Among the clones identified, we focused on TRIP6 (Thyroid Receptor Interacting Protein 6)/ZRP-1 (Zyxin-related Protein 1). TRIP6 has been reported to associate with p130Cas in focal adhesion complexes. We demonstrated that TRIP6 interacted directly with MAGI-1b through binding to the 5th PDZ-binding motif. Ectopic expression of TRIP6 induced invasiveness in the epithelial kidney MDCK and MDCKts-src cells in a PI3-kinase dependant manner, and reverted the E-cadherin-dependent cell-cell aggregation. In this connection, we observed a slight increase in AKT activity in MDCKts-src derivatives transfected with TRIP6 expression vectors, compared to parental cells. The TRIP6Stop473 mutant, lacking the PDZ binding motif was unable to promote cell invasiveness and did not interfere with cell-cell aggregation. The competing peptides corresponding to the C-terminus of TRIP6 or b-catenin promoted invasiveness in TRIP6 Stop473-transfected MDCKts-src cells but not in the parental cell line. These results suggest that TRIP6-induced invasiveness in epithelial cells involves the induction of cell motility by the TRIP6 core molecule, whereas the C-terminus is required to destabilizes E-cadherin junctional complexes by competing with #-catenin for the interaction with MAGI-1b. - 639 -
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Table of content PREFACE ..........
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Preface In 1998, we organized the f
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Acknowledgments This meeting has be
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Group B (15h00 - 17h00) Session V.
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- 11 - Exhibition
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Visit our Expo (in alphabetical ord
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Thursday January 26th, 2006 (Early
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Thursday January 26th, 2006 (Evenin
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Regulation of inflammation and gluc
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Saturday January 28th, 2006 (Early
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Oral presentations (by alphabetical
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4: : Lottin S, Vercoutter-Edouart A
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the B-Raf/mitogen-activated/extrace
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Albertini MC, Accorsi A, Citterio B
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AP-1-dependent gene expression in s
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Jaks and cytokine receptors - an in
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The Role of Met-Gab1 Signalling in
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The mammalian tumor suppressor Scri
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SIGNAL TRANSDUCTION BY STRESS-ACTIV
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Title to be announced Caroline Dive
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Mechanisms of DNA methylation in ma
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Blocking the "4 Integrin-Paxillin I
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Regulation of NF-kB-dependent trans
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The immunomodulator AS101 induces g
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Signaling pathways leading to the a
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Boute, N., Jockers, R. and Issad, T
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Jespsen JS, Sorensen MD and Wengel
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8. Nishikawa, H., Kawai, K., Tanaka
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9) Proteome analysis of rat pancrea
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MicroRNA is an anti-apoptotic facto
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Resveratrol inhibits phorbol ester-
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PI3K Targeting by the Beta-GBP Cyto
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Oncogenic signaling by mutant p53 M
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activity of the Peroxisome Prolifer
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Restauration of SHIP-1 activity in
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Hypoxia Signaling. Impact on Tumor
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HOW ANTITOXIC AND ANTICANCER AGENTS
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Listeria monocytogenes induced Rac1
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Epigenetic Regulation of Stem Cell
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Survey on in vitro cell death induc
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Distinct roles of IRF-3 and IRF-7 i
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Redox-Sensitive Transcription Facto
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Epigenic control of MHC2TA transcri
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Attenuation of MSK1-driven NF-kB ge
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Talking to chromatin: Silencers Sil
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Ubiquitin ligase Smurf1 controls os
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Ambion MicroRNAs (miRNAs) are small
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Organizer: BIOBASE GmbH Date: Janua
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Reliable and efficient gene Knock-d
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Exploring ubiquitin signaling with
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Poster presentations Poster are cla
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Session I : protein domains Poster
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Session II. Receptor signaling and
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Session II : Receptor signaling and
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Session III. Protein kinase cascade
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Session IV. Protein kinase inhibito
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IV. Protein kinase inhibitors: insi
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VI. Proteasome degradation pathways
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VI. Proteasome degradation pathways
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VIII. Inflammation specific signali
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Dr. Jochen Hefl Division of Signal
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