This proposal seeks to identify potential prion proteins within the model plant Arabidopsis thaliana.The term prion was first used to describe the infectious agent causing scrapie, a transmissiblespongiform encephalopathy (TSE) disease, in sheep. Since the n TSE's have been identified in cattle (BSE) and an association between BSE and a variant of a human TSE, Cruetzfeld-Jakob disease (CJD),have been established. The identification of prion-like proteins in Saccharomyces cerevisiae has opened up a new era in t he study of prion proteins.
As well as redefining the term prion to encompass an array of proteins that can behave in an infectious manner, the discovery of prions in yeast allows detailed genetic analysis to be carried out on factors affecting prion stability and propagation using well-established yeast genetic techniques. Furthermore, studies of yeast prions suggest that the prion phenomena may well be widespread throughout nature and that prions have the ability to carry out normal cellular functions. Prionisation domains have been identified and have been shown to be transferable. Using bio-informatics techniques a number of plant proteins that contain putative prion domains have identified. The experimental approach to be taken in this proposal is to transfer these potential plant prion domains and fuse them with green fluorescent protein (GFP) and also use them to replace the prion domain of the known yeast prion protein Sup35. Prions in yeast form aggregates.
We will therefore monitor the aggregation state of GF P in the constructs and observe if they behave like known yeast prions. We will also carry out detailed yeast genetics to investigate whether the plant prion domains can substitute for the natural prion domain of the Sup35 protein. In addition, we will create and screen an Arabidopsis prion domain library constructed from random plant amino acid sequences fused to a yeast selectable marker. Yeast
Field of science
- /natural sciences/chemical sciences/organic chemistry/amines
- /natural sciences/biological sciences/biochemistry/biomolecules/proteins
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