Copper is an essential metal for cellular processes such as detoxification of reactive oxygen species, oxidative phosphorylation, and iron uptake. However, during infection, the host restricts the bioavailability of this micronutrient to the pathogen as a strategy to combat infection. small-RNA-Seq was performed to study the involvement of miRNAs of P. brasiliensis as a response to copper deprivation. Our objective was to characterize the expression profile of miRNAs regulated during copper deprivation in P. brasiliensis and the putative altered processes. For that, P. brasiliensis (ATCC 32069 or Pb18) in the yeast phase were cultured in liquid BHI medium under agitation at 150 rpm at 36C for 72 h. Then, 105 cells/mL were transferred to McVeigh/Morton minimal medium (MMcM) prepared without CuSO4 and supplemented with 50uM of the copper chelator BCS (Sodium betocuproinedisulfonate - Sigma). For the control, the medium was supplemented with 10 uM CuSO4. Treatment and control samples had their RNA extracted with TRIzol from three biological replicates. Samples were pooled and sequenced on Illumina NovaSeq S4 to output approximately 10M 1x50 reads. We identified 14 differentially expressed miRNAs, two of which putatively regulated oxidative stress response, beta-oxidation, glyoxylate cycle, and cell wall remodeling. Our results suggest that metabolic adaptations carried out by P. brasiliensis in copper deprivation are regulated by miRNAs.
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