Glutathione biosynthesis in the yeast pathogens Candida glabrata and Candida albicans: essential in C. glabrata, and essential for virulence in C. albicans.
Redox pathways play a key role in pathogenesis. Glutathione, a central molecule in redox homeostasis in yeasts, is an essential metabolite, but its requirements can be met either from endogenous biosynthesis or from the extracellular milieu. In this report we have examined the importance of glutathione biosynthesis in two major human opportunistic fungal pathogens, Candida albicans and Candida glabrata. As the genome sequence of C. glabrata had suggested the absence of glutathione transporters, we initially investigated exogenous glutathione utilization in C. glabrata by disruption of the MET15 gene, involved in methionine biosynthesis. We observed an organic sulphur auxotrophy in a C. glabrata met15Δ strain; however, unlike its Saccharomyces cerevisiae counterpart, the C. glabrata met15Δ strain was unable to grow on exogenous glutathione. This inability to grow on exogenous glutathione was demonstrated to be due to the lack of a functional glutathione transporter, despite the presence of a functional glutathione degradation machinery (the Dug pathway). In the absence of the ability to obtain glutathione from the extracellular medium, we examined and could demonstrate that γ-glutamyl cysteine synthase, the first enzyme of glutathione biosynthesis, was essential in C. glabrata. Further, although γ-glutamyl cysteine synthase has been reported to be non-essential in C. albicans, we report here for what is believed to be the first time that the enzyme is required for survival in human macrophages in vitro, as well as for virulence in a murine model of disseminated candidiasis. The essentiality of γ-glutamyl cysteine synthase in C. glabrata, and its essentiality for virulence in C. albicans, make the enzyme a strong candidate for antifungal development.
Institute of Microbial Technology (CSIR), Sector 39-A, Chandigarh 160 036, India.
Amit Kumar
AK
Yadav
Institute of Microbial Technology (CSIR), Sector 39-A, Chandigarh 160 036, India.
Prashant Ramesh
PR
Desai
Centre for DNA Fingerprinting and Diagnostics, Building 7, Gruhakalpa 5-4-399/B, Nampally, Hyderabad-500001, India.
Maruti Nandan
MN
Rai
Centre for DNA Fingerprinting and Diagnostics, Building 7, Gruhakalpa 5-4-399/B, Nampally, Hyderabad-500001, India.
Rupinder
R
Kaur
Institute of Microbial Technology (CSIR), Sector 39-A, Chandigarh 160 036, India.
Kaliannan
K
Ganesan
Institute of Microbial Technology (CSIR), Sector 39-A, Chandigarh 160 036, India.
Anand Kumar
AK
Bachhawat
D000818:Animals; D002176:Candida albicans / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D041221:Candida glabrata / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D002460:Cell Line; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D020043:Genes, Essential; D005721:Glutamate-Cysteine Ligase / Q000235:genetics / Q000378:metabolism*; D005978:Glutathione / Q000096:biosynthesis*; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D010084:Oxidation-Reduction; D014774:Virulence
D005656:Fungal Proteins; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione