The Roles of Glycerol in Candida Albicans Biofilm Formation and Invasion

Jigarkumar V. Desai
2018
The fungal pathogen Candida albicans is a leading cause of device-associated and other nosocomial infections. The traits of biofilm formation and invasion into an underlying surface are important for Candida to cause disease. In this dissertation, I describe my work, which reveals a novel role for glycerol in C. albicans biofilm formation and hyphal invasion. Through genomewide expression profiling it was observed that glycerol biosynthetic genes were highly upregulated in biofilms relative to
more » ... he planktonic (suspension) cultures. Consistent with this observation, cells in a biofilm also accumulated higher amounts of glycerol then non-biofilm cells. In order to study the impact of glycerol on biofilm formation I made a deletion mutant, rhr2Δ/Δ, in the gene encoding glycerol-3-phosphatase. Under in vitro conditions, the rhr2Δ/Δ mutant has reduced biofilm biomass and reduced adherence to silicone. The mutant is also severely defective in biofilm formation in the rat venous catheter model of biofilm infection. Surprisingly, genome-wide expression profiling showed that the rhr2Δ/Δ mutant has reduced expression of the cell-surface adhesin genes: ALS1, ALS3, and HWP1, as well as many other genes that are up regulated in biofilms. The role of Rhr2 in adherence and biofilm formation depends on adhesin gene expression as overexpression of any of the adhesin genes restores biofilm formation by rhr2Δ/Δ in vitro and in vivo. Thus, our findings indicate that glycerol plays a regulatory role in biofilm gene expression and that the adhesin genes are among the functional Rhr2-regulated genes. I observed that the functional significance of biofilm glycerol accumulation lies in its ability to generate turgor to drive hyphal invasion. I showed that using an assay for invasive growth into elastic polyacrylamide. Using mathematical and biophysical approaches, I show that C. albicans can generate turgor equivalent to 20 atm in order to invade. Additionally, I show that several mutants with deletions in biofilm and hyphal regulator gen [...]
doi:10.1184/r1/6723776.v1 fatcat:y7l5ktcoibaxti6wqy6zug4dje