Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in Candida glabrata

In this study, we have specifically blocked a key step of sphingolipid (SL) biosynthesis in Candida glabrata by disruption of the orthologs of ScIpt1 and ScSkn1. Based on their close homology with S. cerevisiae counterparts, the proteins are predicted to catalyze the addition of a phosphorylinositol group onto mannosyl inositolphosphoryl ceramide (MIPC) to form mannosyl diinositolphosphoryl ceramide (M(IP)(2)C), which accounts for the majority of complex SL structures in S. cerevisiae membranes. High throughput lipidome analysis confirmed the accumulation of MIPC structures in Delta Cgipt1 and Delta Cgskn1 cells, albeit to lesser extent in the latter. Noticeably, Delta Cgipt1 cells showed an increased susceptibility to azoles; however, Delta Cgskn1 cells showed no significant changes in the drug susceptibility profiles. Interestingly, the azole susceptible phenotype of Delta Cgipt1 cells seems to be independent of the ergosterol content. Delta Cgipt1 cells displayed altered lipid homeostasis, increased membrane fluidity as well as high diffusion of radiolabeled fluconazole (H-3-FLC), which could together influence the azole susceptibility of C. glabrata. Furthermore, in vivo experiments also confirmed compromised virulence of the Delta Cgipt1 strain. Contrarily, specific functions of CgSkn1 remain unclear.

Automated summary

By disrupting the orthologs of ScIpt1 and ScSkn1 in Candida glabrata, we have specifically blocked a key step of sphingolipid biosynthesis and found that it significantly affects the azole susceptibility of the organism.