DOI: 10.1128/spectrum.03874-25 ISSN: 2165-0497

Slt2 kinase of the cell wall integrity pathway is required for Fks2-specific echinocandin resistance in Candida glabrata

Zubayeda Uddin, Saira Tahsin, Klara Valickova, Tengfei Long, Laura Fruhauf De Macedo, Gabrielle Popencuk, Yanan Zhao, Kelley R. Healey

ABSTRACT

Candida glabrata is an opportunistic yeast that causes invasive infections in immunocompromised individuals. Echinocandin antifungals are first-line agents in the treatment of invasive candidiasis and act by inhibiting fungal cell wall synthesis by targeting β-1,3-glucan synthase. Echinocandin resistance is primarily associated with mutations in FKS1 or FKS2 , which encode for the catalytic subunit (Fks1 or Fks2) of the targeted glucan synthase. The cell wall integrity (CWI) pathway, including Slt2 kinase, is known to mediate tolerance to antifungal drugs, but its role in resistance is not completely defined. Here, we observed that disruption of SLT2 abrogated echinocandin resistance due to FKS2 , but not FKS1, mutation in C. glabrata . As expected, reintroduction of plasmid-borne SLT2 restored elevated MICs of fks2 mutants. RT-qPCR revealed that loss of SLT2 led to decreased FKS2 expression levels in wild-type and echinocandin-resistant strains following echinocandin exposure. In conclusion, the CWI pathway and Slt2 comprise a primary control mechanism for FKS2 -mediated echinocandin resistance in C. glabrata .

IMPORTANCE

The higher rates of acquired antifungal resistance exhibited by Candida glabrata merit further understanding of resistance mechanisms including fungal regulation of drug target genes and enzymes. Here, we focused on a fungal tolerance pathway, the cell wall integrity pathway, and its requirement in C. glabrata echinocandin antifungal resistance. Interestingly, we found that targeting the pathway through disruption of a key protein, Slt2, reversed echinocandin resistance within one subset of resistant strains ( fks2 mutants) but not another ( fks1 mutants). Gene expression studies demonstrated the requirement of SLT2 for full FKS2 gene expression following echinocandin treatment. Overall, we discovered that Slt2 is specifically required for Fks2-mediated echinocandin resistance through partial regulation of FKS2 expression. This study provides further insight into the regulation of drug target genes in C. glabrata and provides a possible therapeutic target for echinocandin-resistant infections caused by mutation of FKS2 .

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