Role of nf-kb in the regulation of apoptosis

Линиите свързват координатора с партньорите.

Накратко на български

Цел на проекта

Research objectives and content When cells are exposed to tumor necrosis factor (TNF) two antagonistic pathways are induced: apoptotic and other anti-aptototic. The latter is mediated by the transcription factor NF-KB which is a central mediator for the gene expression induced in cells by pathogens or inflammatory cytokines, and is known to play an important role in development of the immune system. It has been recently shown NF-KB activates an anti apototic response by inducing gene transcription. The aim of our experimental work is to isolate and characterise NF-KB-inducible genes whose protein products have an anti-apoptotic function. Training content (objective, benefit and expected impact) To achieve our objectives, a TNF responsive cell line containing an inducible inhibitor of NF-KB will generated. When the NF-KB inhibitor alpha (a transdominant negative version resistant to signal induced proteolvsis) is induced and the cells exposed to TNF, apotosis will be triggered. In the absence of inhibitor, NF-KB will be induced and transcription of the anti-apoptotic genes will protect the cell from the TNF induced apoptosis. Comparision of the mRNA populations in the presence or absence of IkBa should permit identification of NF-KB inducible genes. Cells expressing the IKBa mutant under negative control of the tetracycline (tet) represor will remain viable in the presence of tet (NF-Kb activated), but will become apoptotic in the absence of tet (NF-KB inhibited). Once an appropriate cell has been identified it will be exposed to TNF and PolyA+ RNA extracted from cells which have expressed the transdominant IKB inhibitor and viable cells in which NF-KB is activated due to repression of the inhibitor gene. mRNAs induced by NF-KB will be isolated by subtractive hybridization using suppression PCR. cDNAs will be labelled and used as hybridization probes to screen a kZAP cDNA lybrary. Additionally cDNAs will be cloned into a conventional vector and sequenced. Known NF-KB induced genes will be discarted and unidentified genes will be further investigated. If those genes products anti-apoptotic activity, they should block apoptosis in TNF treated cells. The study of clAP2 and c) proteins implicated in the modulation of the NF-KB activity and their role on the regulation of its apoptotic activity will be also performed. Links with industry / industrial relevance (22) The host laboratory has colaborations with Pfizer Central Research to search compounds with antiinflammatory activity which inhibit NF-KB.The role of these compounds on anti-apoptotic genes will be tested.

Оригинален текст от CORDIS (на английски).

Участници

• UNIVERSITY OF ST ANDREWS · ST ANDREWSКоординаторОбединеното кралство

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