Project Title and Work Package(s): Structure and role of the Vibrio vulnificus stressosome (WP1, WP2)
Objectives: To: (1) clone, overexpress and purify the components of the V. vulnificus stressosome; (2) perform in vitro assemblies with purified RsbR, RsbS and RsbT and work with participant #2 [UR] to determine structure by cryo-electron microscopy; (3) use gel free proteomics to determine the effects of specific stressosome mutations on the phospho-proteome and total proteome of V. vulnificus when subjected to food related stresses; (4) determine whether the stressosome influences the intracellular pools of the signalling molecule cyclic-diguanylate.
Expected Results: A structural model of the V. vulnificus stressosome including a predicted sensory mechanism. A detailed understanding of the role of the stressosome in modulating the V. vulnificus proteome in response to stress. Elucidation of the downstream signalling events triggered by the stressosome in V. vulnificus.