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Molecular characterization of the Aspergillus fumigatus NCS-1 homologue, NcsA

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Abstract

Here, we characterize the Aspergillus fumigatus homologue ncsA Neuronal Calcium Sensor. We showed that ncsA is not an essential gene and ∆ncsA growth was decreased in the presence of EGTA and SDS. Furthermore, the ∆ncsA mutant is more resistant to calcium chloride. NcsA:mRFP localizes to the cytoplasm and its cellular localization is not affected by the cellular response to either calcium chloride or EGTA. The ∆ncsA mutant strain is more sensitive to voriconazole, itraconazole, and amphotericin. Polar growth in the ΔncsA mutant was also considerably more affected by lovastatin than in the wild type strain. The Spitzenkörper can be visualized in both strains and although the vacuolar system does not seem to be very different, there is an increase in the staining intensity on the germling surface of the ∆ncsA strain. NcsA promotes pmcA and pmcB expression and therefore there is a reduced expression of these ion pumps in the ΔncsA mutant background, and also of other genes involved in the response to calcium in A. fumigatus. The ncsA inactivation mutation is not causing loss of virulence in a low dose murine infection when compared to the corresponding wild type strain.

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Acknowledgements

This research was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brazil, and John Simon Guggenheim Memorial Foundation, USA (to GHG); and the Deutsche Forschungsgemeinschaft (SPP1160), Germany (AAB). We would like to acknowledge Natalie Fedorova for helping in the identification of A. fumigatus proteins with EF-hands domains, the two anonymous reviewers for their comments, and the Laboratory of Confocal Microscopy from the Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Brazil, for the use of the confocal microscope.

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  1. Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. do Café S/N, CEP 14040-903, Ribeirão Preto, São Paulo, Brazil

    André Oliveira Mota Júnior, Iran Malavazi, Frederico Marianetti Soriani, Marcela Savoldi, Márcia Eliana da Silva Ferreira & Gustavo Henrique Goldman

  2. Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology-Hans-Knoell-Institute (HKI), Jena, Germany

    Thorsten Heinekamp & Axel A. Brakhage

  3. Experimental Biology Infection and Drug Testing Group, Leibniz Institute for Natural Product Research and Infection Biology-Hans-Knoell-Institute (HKI), Jena, Germany

    Ilse Jacobsen

  4. Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, São Paulo, Brazil

    Maria Helena S. Goldman

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  1. André Oliveira Mota Júnior
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  2. Iran Malavazi
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  9. Márcia Eliana da Silva Ferreira
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Correspondence to Gustavo Henrique Goldman.

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Communicated by J. Perez-Martin.

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Mota Júnior, A.O., Malavazi, I., Soriani, F.M. et al. Molecular characterization of the Aspergillus fumigatus NCS-1 homologue, NcsA. Mol Genet Genomics 280, 483–495 (2008). https://doi.org/10.1007/s00438-008-0381-y

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