Functional characterization of an Aspergillus fumigatus calcium transporter (PmcA) that is essential for fungal infection

• Published in: PloS one 2012-05, Vol.7 (5), p.e37591-e37591
• Main Authors: Dinamarco, Taísa Magnani, Freitas, Fernanda Zanolli, Almeida, Ricardo S, Brown, Neil Andrew, dos Reis, Thaila Fernanda, Ramalho, Leandra Naira Zambelli, Savoldi, Marcela, Goldman, Maria Helena S, Bertolini, Maria Célia, Goldman, Gustavo Henrique
• Format: Article
• Language: English
• Subjects: ABC transporters; Allergens; Alveoli; Animal models; Animals; Antifungal agents; Aspergillosis; Aspergillus fumigatus; Aspergillus fumigatus - enzymology; Aspergillus fumigatus - growth & development; Aspergillus fumigatus - pathogenicity; Aspergillus nidulans; Binding; Biology; Bronchoalveolar Lavage - methods; Ca2+-transporting ATPase; Calcineurin; Calcium; Calcium Signaling - physiology; Candida albicans; Conditional mutant; DNA Primers - genetics; Efficiency; Electrophoretic Mobility Shift Assay; Fungal Proteins - genetics; Fungal Proteins - metabolism; Fungicides; Gene Expression Regulation, Fungal - genetics; Gene Expression Regulation, Fungal - physiology; Gene sequencing; Genes; Genetic Vectors - genetics; Genomics; Health aspects; Manganese; Metabolism; Mice; Morphogenesis; Morphology; mRNA; Mutants; Mycoses; Opportunist infection; Paracoccidioides brasiliensis; Plasma Membrane Calcium-Transporting ATPases - genetics; Plasma Membrane Calcium-Transporting ATPases - metabolism; Proteins; Pulmonary Aspergillosis - enzymology; Pulmonary Aspergillosis - microbiology; Real-Time Polymerase Chain Reaction; Regulatory sequences; Response Elements - genetics; Reverse Transcriptase Polymerase Chain Reaction; RNA; Saccharomyces cerevisiae; Salts; Signal transduction; Transcription factors; Transporter; Virulence; Virulence - genetics; Yeast
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0037591

Aspergillus fumigatus is a primary and opportunistic pathogen, as well as a major allergen, of mammals. The Ca(+2)-calcineurin pathway affects virulence, morphogenesis and antifungal drug action in A. fumigatus. Here, we investigated three components of the A. fumigatus Ca(+2)-calcineurin pathway, pmcA,-B, and -C, which encode calcium transporters. We demonstrated that CrzA can directly control the mRNA accumulation of the pmcA-C genes by binding to their promoter regions. CrzA-binding experiments suggested that the 5'-CACAGCCAC-3' and 5'-CCCTGCCCC-3' sequences upstream of pmcA and pmcC genes, respectively, are possible calcineurin-dependent response elements (CDREs)-like consensus motifs. Null mutants were constructed for pmcA and -B and a conditional mutant for pmcC demonstrating pmcC is an essential gene. The ΔpmcA and ΔpmcB mutants were more sensitive to calcium and resistant to manganese and cyclosporin was able to modulate the sensitivity or resistance of these mutants to these salts, supporting the interaction between calcineurin and the function of these transporters. The pmcA-C genes have decreased mRNA abundance into the alveoli in the ΔcalA and ΔcrzA mutant strains. However, only the A. fumigatus ΔpmcA was avirulent in the murine model of invasive pulmonary aspergillosis.