Zrg1, a cryptococcal protein associated with regulation of growth in nutrient deprivation conditions

The influence of amoeba metal homeostasis on antifungal activity against Cryptococcus gattii

“…Trophozoites of A. castellanii Neff strain (kindly provided by Dr Allan Jefferson Guimarães - Universidade Federal Fluminense) were axenically cultured at 25 °C in peptone-yeast extract glucose (PYG) medium (20 g/L peptone, 2 g/L yeast extract, 0.1 M glucose, 4 mM MgSO 4 , 3.4 mM sodium citrate, 0.9 mM Fe(NH 4 ) 2 (SO4) 2 , 1.3 mM Na 2 HPO 4 , and 2 mM K 2 HPO 4 , pH 6.5) supplemented with 20 U/ml penicillin, 20 U/ml streptomycin, and 20 U/ml chloramphenicol in 12-well cell culture plates. The C. gattii strain R265 (WT) and the null mutant strains for the ZIP1 gene ( zip1Δ ) ( Schneider et al, 2015 ), ZIP3 gene ( zip3Δ ) ( Garcia et al, 2020 ) and ZRG1 gene ( zrg1Δ ) ( Diehl et al, 2021 ) were used in this work. The yeast strains were routinely cultured in YPD medium (2% glucose, 2% peptone, and 1% yeast extract) and incubated in an orbital shaker (200 rpm) at 30 °C overnight.…”

“…Zinc is an important transition metal for virtually all living cells ( Cuajungco et al, 2021 ). Our group previously showed that zinc levels regulate the expression of several genes in the fungal pathogen C. gattii ( Schneider et al., 2012 ; Diehl et al, 2021 ). Among such genes are the metal transporter coding genes ZIP1 (CNBG_6066) and ZIP3 (CNBG_5361).…”

“…ZIP3 null mutants displayed altered manganese homeostasis and hypersensitivity to oxidative stress ( Garcia et al, 2020 ). Zinc availability also regulates the expression of a gene ( ZRG1 - CNBG_1485) that controls proper autophagy in C. gattii ( Diehl et al, 2021 ). Due to its essentiality, zinc is a target of nutritional immunity.…”