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Identifying components of the yeast ERAD pathway and their interaction with a toxic substrate

Reference	C13717
Principal Investigator / Supervisor	Professor Lynne Roberts
Co-Investigators / Co-Supervisors	Professor John Davey, Professor Michael Lord, Professor Colin Stirling
Institution	University of Warwick
Department	Biological Sciences
Funding type	Research
Value (£)	178,984
Status	Completed
Type	Research Grant
Start date	01/02/2001
End date	29/02/2004
Duration	37 months

Abstract

Proteins in the ER lumen that fail to fold or assemble correctly are degraded by the ER-associated protein degradation pathway (ERAD). The toxic ricin A chain (RTA) also utilises ERAD to translocate into the cytosol from the ER. Here we propose to generate RTA from an inactive precursor within the ER lumen of yeast. This will be lethal for the cell and forms the basis of a novel screen to identify mutants unable to export RTA by the ERAD pathway. Such mutants should survive because the RTA generated will remain trapped within the ER. ER lumenal and translocation channel components that interact with RTA during export will also be identified by chemical cross-linking and by trapping translocating intermediates of RTA-dihydrofolate reductase (DHFR) fusions in the presence of DHFR substrate analogues. The topology of RTA during its dislocation from the ER will also be investigated.

Summary

unavailable

Committee	Closed Committee - Biochemistry & Cell Biology (BCB)
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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