Development and application of fluorescent proteins for endogenous imaging of lowly-expressed proteins and elucidating the role of the G-protein coupled receptor, Gpr1, in the virulence of Candida albicans

Infections caused by Candida albicans are becoming harder to eradicate due to increasing resistance against the widely used antifungals. In addition to the increasing resistance, the population of immunocompromised people (e.g. the elderly, neonates, etc.), which are extremely susceptible, is expanding. Therefore, novel therapies have to be discovered to treat and prevent fungal infections if we do not want to lose the arm's race between antifungals and resistance mechanisms. One of the key virulence factors of C. albicans is biofilm formation. Biofilms are microbial communities embedded in an extracellular matrix and attached to a biotic or abiotic surface. Key processes in biofilm formation are morphogenesis (the transition from a round to an elongated form) and adhesion. These processes are both regulated by the Protein Kinase A pathway (PKA). A biosensor will be developed for Candida albicans using existing PKA biosensors as scaffolds. By incorporating a photoswitchable FRET pair into this scaffold we will achieve a robust biosensor capable of dealing with the high autofluorescence of Candida cells. On top of that the biosensor will allow for a quick and easy super resolution readout of PKA activity. Using this biosensor, we will set up a screening method to discover inhibitors that specifically target the G-protein coupled receptor system of the PKA pathway. The hit compounds acquired from this screening method may form the basis for novel antifungals.

Publication year:2021

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