Objective
Research objectives and content
Preliminary results show that the Chediak-Higashi syndrome protein (lyst) is implicated in protein sorting in the endosomes. We plan to analyze the role of lyst in endosomal protein sorting by identification of lyst domains implicated in this process and lyst binding proteins also implicated in endosomal protein sorting. Objectives:
1.- Confirmation of the direct implication of lyst in endosomal protein sorting in fibroblasts. First, we will confirm that the CHS mutation of lyst is affecting endosomal protein sorting in fibroblasts. Transient expression of suitable plasmid constructs carrying the wild type lyst cDNA will be performed in defective CHS fibroblasts to analyze recovery of normal phenotype.
2.- Identification of lyst domains implicated in endosomal protein sorting. The functional study of different lyst domains will be performed by transfecting full length or mutated plasmid constructs containing truncated versions of lyst into CHS defective fibroblasts, providing the minimal lyst sequence necessary for correct endosomal protein sorting. 2.3.- Identification of lyst binding proteins in fibroblasts. The identification of lyst binding proteins will provide crucial information regarding its function in the endosomes, it will either implicate known proteins (may be, coat proteins) or identify novel proteins directly or indirectly involved in this process. This objective will be approached by three different strategies:
a) Two hybrid system b) Binding to recombinant lyst protein c) Co-immunoprecipitations
Training content (objective, benefit and expected impact) The recent finding that the protein defective in CHS is involved in endosomal sorting opens novel possibilities to analyze this particular step of intracellular transport. This project will provide essential information about a fundamental cell mechanism which is currently poorly understood: the sorting of proteins in endosomes. Because this defect results in a severe immunodeficiency, this project will also contribute to the understanding of how basic mechanisms of intracellular transport contribute to develop an efficient immune response.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences biological sciences biochemistry biomolecules proteins
- medical and health sciences basic medicine immunology
- natural sciences biological sciences genetics mutation
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Multi-annual funding programmes that define the EU’s priorities for research and innovation.
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Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
Coordinator
75248 PARIS
France
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