Towards a temperature-sensitive proteome: developing a Drosophila-friendly degron

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Here we propose to develop a new technology that enables biologists to turn activity of genes on and off simply by changing the temperature, by targeting their products for degradation only at higher culture temperatures. This technology is based on well-established biological mechanisms employed by the cell to achieve regulated removal of unwanted proteins. This regulated degradation is mediated by the attachment of degradation signals to the target. Some proteins (degrons), identified in yeast, can induce attachment of these signals to themselves when their structure is disrupted at higher temperatures, thus inducing degradation of themselves and any fusion proteins that they are part of. We and others have successfully shown proof-of-principle that the system can be used in a more complex model organism, Drosophila (fruitfly). However, the temperatures at which degradation of the target is induced in yeast are not well-tolerated by Drosophila, or by other 'cold-blooded' organisms. Therefore, we propose to isolate novel variants of the yeast degron signal, that work better in the normal temperature ranges of Drosophila and of other model organisms widely used by biologists, and to test these in cultured Drosophila cells and in genetically manipulated flies.