Visit our Expo - Redox and Inflammation signaling 2012

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Session II : Receptor signaling and G proteins Poster II, 17 Functional overlapping between caspase cleavage site and CBL binding sequence in the juxtamembrane domain of the MET receptor. Julien DEHEUNINCK, Catherine LEROY, Zongling JI, Bénédicte FOVEAU, Vincent VILLERET, David TULASNE and Véronique FAFEUR CNRS UMR 8117, Institut de Biologie de Lille, Institut Pasteur de Lille, B.P.447, 59021 Lille, FRANCE. E-mail : veronique.fafeur@ibl.fr Hepatocyte growth factor/ scatter factor (HGF/SF) binds and activates the MET tyrosine kinase receptor to transduce cell survival. Nonetheless, we previously demonstrated that stress stimuli cause a caspase-dependent cleavage in the MET juxtamembrane domain, which converts MET into a pro-apoptotic p40 MET fragment. The caspase 3 cleavage occurs at aspartic acid residue D1000. This residue is juxtaposed to tyrosine Y1001, which is rather involved in binding CBL upon MET activation by its ligand, allowing MET trafficking and recycling. We wished to examine the functional relationships between caspase-dependent cleavage and CBL binding at the ESVDY sequence of the juxtamembrane domain of MET. We first established that the caspase cleavage- and CBL binding- sites overlapped, since a MET D1000N mutant lost the ability both to generate p40 MET and to recruit CBL. We then individually mutated all amino acids of the ESVDY sequence of MET constituting the caspase and CBL sites. While a MET V999A mutant was selectively defective in generating p40 MET, a MET Y1001F mutant was selectively defective in recruiting CBL, demonstrating that caspase-dependent cleavage and CBL recruitment are dissociable mechanisms. We further examined the consequence of MET phosphorylation at CBL binding site on the caspase-dependent cleavage. Using in vitro MET phosphorylation and dephosphorylation, we demonstrated that generation of p40 MET is unfavored when MET is phosphorylated, and vice versa. These data were strengthened by modeling studies using the MET peptide sequence ESVDYR and the known structure of caspase 3 or CBL in complex with specific peptide sequences. These studies show that a phosphorylated Y1001 is not compatible with specific association of caspase-3 to MET and also exclude simultaneous fixation of both caspase 3 and CBL to MET. In conclusion, we demonstrated that the caspase-dependent cleavage of MET and binding of CBL to the activated MET receptor are dissociable and incompatible mechanisms. As a consequence, we propose that HGF/SF can protect MET against caspase-dependent cleavage by activating autophosphorylation of the receptor. - 144 -
Session II : Receptor signaling and G proteins Poster II, 18 Identification of PAR1-G protein signalling pathways involved in thrombin-induced CCL2 release Xiaoling Deng, Paul F Mercer, Geoffrey J Laurent and Rachel C Chambers. Centre for Respiratory Research, University College London, 5 University Street, WC1E 6JJ, London, England. E-mail: x.deng@ucl.ac.uk Introduction: In experimental models of lung injury, activation of proteinase-activated receptor-1 (PAR1) by coagulation proteinases is critical in driving both the inflammatory and fibrotic response. PAR1 exerts its pluripotent cellular effects by concomitant activation of G"i/o, G"q and G"12/13. We have previously shown that protection from lung inflammation and fibrosis in PAR1 deficient (PAR1 KO) mice is accompanied by reduced lung levels of the potent thrombin-inducible protein CCL2 (MCP-1/JE). The aim of this study was to examine the signalling pathways by which PAR1 activation induces the release of this chemokine by cultured mouse lung fibroblasts. Methods and Results: Wild type mouse lung fibroblasts (MLFs) were stimulated with thrombin, TFLLR-NH2 and RLLFT-NH2 (PAR1 peptide agonist and control peptide respectively), and CCL2 protein release into cell culture supernatants was assessed by ELISA. Both thrombin and TFLLR induced CCL2 release by MLFs in a time- and dose-dependent manner. No response was obtained with the control peptide RLLFT-NH2 or in PAR1 KO fibroblasts. The PAR1 specific antagonist RWJ-58259 (1µM) completely blocked thrombin induced-CCL2 release. In order to investigate potential signalling pathways involved the effects of pertussis toxin (PTX, G"i/o inhibitor), Ro-318425 (Protein Kinase C (PKC) inhibitor and U0126 (MEK1/2 inhibitor) on PAR1 activation induced-CCL2 release was also assessed. PTX inhibition of G"i/o had no effect on CCL2 release induced by thrombin, whereas Ro-318425 inhibition of PKC reduced CCL2 release by 49±6%, and U0126 inhibition of MEK1/2 reduced CCL2 release by 53±8% (all p
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- 1 - Luxembourg, January 25th to 2
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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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Cross-talk between angiotensin II a
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Multiple role of histamine H 1 rece
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PI3K Targeting by the Beta-GBP Cyto
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D.P. Chopra, R.E. Menard, J. Janusz
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[2] Meijer, L., Flajolet, M. and Gr
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[4] Niemand, C., Nimmesgern, A., Ha
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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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5. Sokolov EI, Zykov KA, Pukhalsky
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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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Session III : Protein kinase cascad
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Session IV. Protein kinase inhibito
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IV. Protein kinase inhibitors: insi
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IV. Protein kinase inhibitors: insi
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V. Phosphatases as key cell signali
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VI. Proteasome degradation pathways
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VI. Proteasome degradation pathways
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Session VII. Stem cell specific cel
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VIII. Inflammation specific signali
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Session IX. Novel compounds targeti
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Session X : Cell death in cancer -
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Session X : Cell death in cancer Po
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Session XI : Cell death and cardiov
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Session XI: Cell death and cardiova
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Session XII : Cell death and neurod
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Session XIII : Cell signaling pathw
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Session XIV : Transcriptional and t
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Session XV : Reactive oxygen specie
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Session XVI : Chemopreventive agent
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Session XVII : Cell signaling in he
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Dr. Nassera Aouali L-1526 Luxembour
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Dr. Giordano Bianchi Clinic of inte
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Mrs. Isabel Burghardt Laboratory of
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Pr. Czeslaw Cierniewski 92-215 Lodz
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Mr. Igotz Delgado Biochemistry 4894
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Mrs. Anissa Fergani INSERM U-692 67
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Dr. Mark Ginsberg Mail code 0726 92
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Dr. Jochen Hefl Division of Signal
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Mr. Jan Jepsen 2100 Copenhagen DNK
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Pr. Young Min Kim Division of Biolo
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Mr. Christoph Lahtz AG Tumorgenetik
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Pr. Etta Livneh 84105 Beer Sheva IS
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Mrs. Rasa Merzvinskyte Department o
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Mr. Gustav Nilsonne Department of L
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Mrs. Christel Péqueux Tumour and D
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Dr. Gabriella Regis Tumor Immunolog
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Dr. Carsten Scheller 97078 Wuerzbur
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Mr. Yoo-Cheol Song Laboratory of Gy
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Mrs. Jessica Wagener 40225 Duesseld
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Pr. Moncef ZOUALI Centre Viggo Pete
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