Chemical Genetic Profiling and Characterization of Small-molecule Compounds That Affect the Biosynthesis of Unsaturated Fatty Acids in Candida albicans

Prostaglandins are C20 fatty acid metabolites with diverse biological functions. In mammalian cells, prostaglandins are produced from arachidonic acid (AA) via cyclooxygenases (COX1 and COX2). Although fungi do not possess cyclooxygenase homologues, several pathogenic species are able to produce prostaglandins from host-derived arachidonic acid. In this study, we characterized the prostaglandin profile of the emerging human pathogen Candida parapsilosis with HPLC-MS and compared it to that of C. albicans. We found that both species synthesized prostaglandins (mainly PGD 2 and PGE 2 ) from exogenous AA. Furthermore, as OLE2 has been associated with prostaglandin synthesis in C. albicans, we generated homozygous OLE2 deletion mutants in C. parapsilosis and examined their PGE 2 production. However, the PGE 2 production of the OLE2 KO strain was similar to that of wild type (WT), indicating that OLE2 is not required for prostaglandin synthesis in C. parapsilosis. Interestingly, analyses of the fatty acid composition of WT and OLE2 KO cells by gas chromatography (GC) highlighted the accumulation of palmitoleic and oleic acid in the OLE2 deletion mutant. The OLE2 KO cells were killed more efficiently by human monocytes-derived macrophages (MDMs) as well as induced higher interleukin-10 (IL-10) secretion, indicating that OLE2 affects the virulence of C. parapsilosis. Taken together, these results contribute to the better understanding of fatty acid biosynthesis pathways in C. parapsilosis.

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“…17,22 In accordance with these findings, our results further support the role of fatty acid homeostasis in the virulence of pathogenic fungi.…”

Section: Discussionsupporting

confidence: 80%

“…17,21 Importantly, Ole1 plays a crucial role in virulence in both species, as demonstrated by in vivo murine infection models. 17,22 However, while OLE2 has been associated with prostaglandin synthesis in C. albicans, the role of OLE2 in C. parapsilosis is unknown.…”

Section: Introductionmentioning

confidence: 99%

Candida parapsilosis produces prostaglandins from exogenous arachidonic acid and OLE2 is not required for their synthesis

Grózer

Tóth

et al. 2015

Virulence

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“…L. monocytogenes strains used in this study include wild-type (WT) strain 10403S; DP-L3903; the ⌬hly mutant, with a clean deletion of the gene encoding listeriolysin O; the cld-2 mutant, which harbors a Tn917 insertion in the gene encoding the E3 subunit of the BKD complex (25); and two transductants, BKD-1 and BKD-2, which are WT strains independently transduced with the cld-2 transposon allele. Transduction was accomplished by infecting WT bacteria with U153 bacteriophage stock packaged with genomic DNA from the cld-2 mutant and selecting for erythromycin resistance, a marker present on Tn917.…”

Section: Methodsmentioning

confidence: 99%

Branched-Chain Fatty Acids Promote Listeria monocytogenes Intracellular Infection and Virulence

Sun

O’Riordan

2010

Infect Immun

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Anteiso-branched-chain fatty acids (BCFA) represent the dominant group of membrane fatty acids and have been established as crucial determinants in resistance against environmental stresses in Listeria monocytogenes, a facultative intracellular pathogen. Here, we investigate the role of anteiso-BCFA in L. monocytogenes virulence by using mutants deficient in branched-chain alpha-keto acid dehydrogenase (BKD), an enzyme complex involved in the synthesis of BCFA. In tissue culture models of infection, anteiso-BCFA contributed to intracellular growth and survival in macrophages and significantly enhanced plaque formation upon prolonged infection in L2 fibroblasts. The intracellular defects observed could be attributed partially to insufficient listeriolysin O (LLO) production, indicating a requirement for anteiso-BCFA in regulating virulence factor production. In a murine model of infection, the BKD-deficient mutant was highly attenuated, further emphasizing the importance of BKD-mediated metabolism in L. monocytogenes virulence. This study demonstrates an underappreciated role for BCFA in bacterial pathogenesis, which may provide insight into the development and application of antimicrobial agents.

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“…To make matters worse, the number of effective antifungal drugs is limited, and resistant strains to frequently-used drugs, including echinocandin, have been emerging in clinical settings (Perlin, 2007). In this background, offering novel drug targets and inhibitors or ligands will help in antifungal drug development (Jiang et al, 2008;Xu et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Target validation and ligand development for a pathogenic fungal profilin, using a knock‐down strain of pathogenic yeast Candida glabrata and structure‐based ligand design

Ueno

Tsutsumi

Chibana

2010

Yeast

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The emergence of antifungal drug resistance is triggering vigorous searches for novel antifungal targets and lead compounds. In this study, we focused on fungal profilin, which is a small actin control protein sharing limited homology to human profilin. To validate its potentiality as a target, a profilin-conditional mutant of the pathogenic yeast Candida glabrata was constructed, using a regulatable Tet promoter, and its growth was monitored in vitro. Repression of profilin expression led to severe growth defect, demonstrating the potential of this protein as a novel antifungal target. Next, novel peptides binding to the active interface of profilin were designed by computer simulation. ELISA analysis showed that these peptides did bind to the wild-type profilin but bound less strongly to a profilin with amino acid substitutions at the active interface. Hence, we show here that profilin is a potential antifungal target and offer novel peptide ligands.

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Chemical Genetic Profiling and Characterization of Small-molecule Compounds That Affect the Biosynthesis of Unsaturated Fatty Acids in Candida albicans

Cited by 69 publications

References 34 publications

Candida parapsilosis produces prostaglandins from exogenous arachidonic acid and OLE2 is not required for their synthesis

Candida parapsilosis produces prostaglandins from exogenous arachidonic acid and OLE2 is not required for their synthesis

Branched-Chain Fatty Acids Promote Listeria monocytogenes Intracellular Infection and Virulence

Target validation and ligand development for a pathogenic fungal profilin, using a knock‐down strain of pathogenic yeast Candida glabrata and structure‐based ligand design

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