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Molecular analysis of interactions between the resistance signalling proteins EDS1 and PAD4 and the senescence regulator SAG101

Reference	BB/C50135X/1
Principal Investigator / Supervisor	Dr Bart Feys
Co-Investigators / Co-Supervisors
Institution	Imperial College London
Department	Biological Sciences
Funding type	Research
Value (£)	261,856
Status	Completed
Type	Research Grant
Start date	16/11/2004
End date	15/03/2008
Duration	40 months

Abstract

The Arabidopsis proteins EDS1 and PAD4 are central regulators of both R gene mediated and basal resistance, and are required for the accumulation of the defence signalling molecule salicylic acid during pathogen interactions. Both proteins are known to form a complex in plant cells that is under positive feedback control to amplify the defence response. Affinity purification of EDS1-containing protein complexes from plant cells, followed by mass-spectrometric analysis, has recently identified a senescence regulator, SAG101, as an in planta EDS1-interacting protein. This proposal will look at genetic interactions between disease resistance and senescence through the isolation of knockout mutants and double/triple mutant combinations. This genetic framework will allow a further analysis of complex composition and dynamics within the EDS1-PAD4-SAG101 family of proteins using size-exclusion chromatography. This family of proteins has sequence motifs that are indicative of a lipase-like, or in more general terms an acyl hydrolytic activity. As a continuation of previous analyses, we will test whether the predicted catalytic residues in EDS1 and PAD4 are required for in planta function through site directed mutagenesis and complementation of double and triple mutant backgrounds. In a complementary approach, we aim to provide insights into EDS1 and PAD4 biochemical function through a structural biology approach. This will involve co-expression in E. coli of EDS1-containing complexes that we know to be present in plant cells. Various combinations of vectors and host strains will be tested to define optimal conditions for large scale expression of soluble protein as a first step towards structural characterisation of this important class of signalling proteins.

Summary

unavailable

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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