IFCPAR AR (ENGLISH) for CD - CEFIPRA

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CEFIPRA Centre Franco-Indien pour la Promotion de la Recherche Avancée Duration: Three years (December, 2008 to November, 2011) Objectives i) Establish the list of all genes (genome-wide coverage) whose siRNA-mediated downmodulation affects the endocytosis and retrograde transport of Shiga toxin (defined as Shiga siRNAome) in a Drosophila cell line, and validate targets in a human cell line ii) Comparison between the siRNAomes of Shiga toxin and GPI-GFP and with the published literature on similar experiments with SV40 virus, the collaborators hope to establish common principles and specific differences for the clathrin-independent cellular uptake process of these marker proteins iii) Establish the list of genes (genome-wide coverage) whose siRNA-mediated downmodulation affects the endocytosis and retrograde transport of ricin (defined as ricin siRNAome) iv) Compare siRNAomes of Shiga toxin and ricin and establish common principles and specific differences for the retrograde transport of these protein toxins v) Through both: a) identification of key regulators of Shiga toxin and ricin entry into cells, and b) lay the groundwork for the development of toxin inhibitors (small molecules) that would respond to medical needs in infectious disease (hemolytic-uremic syndrome) and protection against bioterrorism Accomplishments i) Screening of more than 600 tagged proteins for their localization on tubular endocytic membranes invaginations induced by Shiga toxin; identification of 10 hits = cytosolic, plasma membrane, cytoskeleton, and Rab proteins ii) Identification of cellular proteins that like Shiga toxin induce plasma membrane invaginations iii) Analysis of glycosphingolipid clustering by Shiga toxin and cellular proteins by fluorescence anisotropy measurements; labeling of cells with synthetic glycolipids to obtain conditions optimum for Shigaintoxication iv) Development of proteomics and lipidomics approaches on transport intermediates of clathrin-independent endocytosis Research papers published: One Papers presented in conference: 5 Project 3803-2 siRNAome OF TOXIN ENDOCYTOSIS AND RETROGRADE TRANSPORT Research Activities 2010-11 Life and Health Sciences Prof. Satyajit Jitu Mayor National Centre for Biological Sciences Bangalore Prof. Ludger Johannes Institut Curie UMR 144 Curie/CNRS Paris Co-localization between a surface cargo in protein induced tabules 37
38 Life and Health Sciences Dr. Shyamala Mani Indian Institute of Science Bangalore Prof. Pierre Gressens INSERM U 676 IFR 02 & IFRH Hôpital Robert Debré(AP-HP) Paris Project 3803-3 NEURAL DIFFERENTIATION OF EMBRYONIC STEM CELLS IFCPAR Indo-French Centre for the Promotion of Advanced Research Duration: Three years (January, 2009 to December, 2011) Objectives i) To delineate the role of proneural transcription factors in neuronal subtype determination in human and mouse embryonic stem cells ii) To determine the role of extrinsic factors on neural differentiation Accomplishments i) To fulfil the goal of the first objective the collaborators created a mouse embryonic stem cell line that expresses the proneural transcription factor, that the collaborators can turn off and on at different time points during neural differentiation of the ES cells ii) Assays have been standardized that will allow collaborators to differentiate mouse ES cells into neuronal lineage and to look at the effect of the proneural transcription factors on the subtype of neurons that are being produced iii) To address the second objective extrinsic factors that are important for granule neuron differentiation have been looked into and how these factors interact with intrinsic determinants of neuronal subtype has been studied iv) The sonic hedgehog signalling pathway has been characterised during human cerebellar development. Cerebellar development in cases of babies born prematurely has been characterised to understand how the signalling pathways have been perturbed. Research papers published: Five Poster: 2
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