The Role of RodA-Conserved Cysteine Residues in the Aspergillus fumigatus Conidial Surface Organization

Institut National de la Santé et de la Recherche Médicale, Centre de Recherché des Cordeliers, Sorbonne Université, Université de Paris, 75006 Paris, France

Molecular Mycology Unit, Institut Pasteur, CNRS-UMR-2000, 10098 Paris, France

⁴

School of Medical Sciences and Sydney Nano, University of Sydney, Sydney, New South Wales (NSW) 2006, Australia

⁵

Biological NMR Technological Platform, Institut Pasteur, CNRS-UMR-3528, 75015 Paris, France

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Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, 07745 Jena, Germany

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Medical Research Council Centre for Medical Mycology (MRC CMM), University of Exeter, Exeter EX4 4QD, UK

⁸

Institute of Microbiology, Friedrich Schiller University, 07745 Jena, Germany

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

^‡

Current address: Division of Immunology, Boston Children’s Hospital, Boston, MA 021115, USA.

^§

Current address: School of Medicine, University of Crete, 71003 Heraklion, Crete, Greece.

J. Fungi 2020, 6(3), 151; https://doi.org/10.3390/jof6030151

Received: 9 July 2020 / Revised: 21 August 2020 / Accepted: 24 August 2020 / Published: 26 August 2020

(This article belongs to the Special Issue Innate Immunity in Fungal Infections)

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Abstract

Immune inertness of Aspergillusfumigatus conidia is attributed to its surface rodlet-layer made up of RodAp, characterized by eight conserved cysteine residues forming four disulfide bonds. Earlier, we showed that the conserved cysteine residue point (ccrp) mutations result in conidia devoid of the rodlet layer. Here, we extended our study comparing the surface organization and immunoreactivity of conidia carrying ccrp-mutations with the RODA deletion mutant (∆rodA). Western blot analysis using anti-RodAp antibodies indicated the absence of RodAp in the cytoplasm of ccrp-mutant conidia. Immunolabeling revealed differential reactivity to conidial surface glucans, the ccrp-mutant conidia preferentially binding to α-(1,3)-glucan, ∆rodA conidia selectively bound to β-(1,3)-glucan; the parental strain conidia showed negative labeling. However, permeability of ccrp-mutants and ∆rodA was similar to the parental strain conidia. Proteomic analyses of the conidial surface exposed proteins of the ccrp-mutants showed more similarities with the parental strain, but were significantly different from the ∆rodA. Ccrp-mutant conidia were less immunostimulatory compared to ∆rodA conidia. Our data suggest that (i) the conserved cysteine residues are essential for the trafficking of RodAp and the organization of the rodlet layer on the conidial surface, and (ii) targeted point mutation could be an alternative approach to study the role of fungal cell-wall genes in host–fungal interaction. View Full-Text

Keywords: Aspergillus fumigatus; host–fungal interaction; conidial surface; RodAp; hydrophobin; disulfide bonds; immunomodulation

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MDPI and ACS Style

Valsecchi, I.; Stephen-Victor, E.; Wong, S.S.W.; Karnam, A.; Sunde, M.; Guijarro, J.I.; Rodríguez de Francisco, B.; Krüger, T.; Kniemeyer, O.; Brown, G.D.; Willment, J.A.; Latgé, J.-P.; Brakhage, A.A.; Bayry, J.; Aimanianda, V. The Role of RodA-Conserved Cysteine Residues in the Aspergillus fumigatus Conidial Surface Organization. J. Fungi 2020, 6, 151. https://doi.org/10.3390/jof6030151

AMA Style

Valsecchi I, Stephen-Victor E, Wong SSW, Karnam A, Sunde M, Guijarro JI, Rodríguez de Francisco B, Krüger T, Kniemeyer O, Brown GD, Willment JA, Latgé J-P, Brakhage AA, Bayry J, Aimanianda V. The Role of RodA-Conserved Cysteine Residues in the Aspergillus fumigatus Conidial Surface Organization. Journal of Fungi. 2020; 6(3):151. https://doi.org/10.3390/jof6030151

Chicago/Turabian Style

Valsecchi, Isabel, Emmanuel Stephen-Victor, Sarah Sze Wah Wong, Anupama Karnam, Margaret Sunde, J. Iñaki Guijarro, Borja Rodríguez de Francisco, Thomas Krüger, Olaf Kniemeyer, Gordon D. Brown, Janet A. Willment, Jean-Paul Latgé, Axel A. Brakhage, Jagadeesh Bayry, and Vishukumar Aimanianda. 2020. "The Role of RodA-Conserved Cysteine Residues in the Aspergillus fumigatus Conidial Surface Organization" Journal of Fungi 6, no. 3: 151. https://doi.org/10.3390/jof6030151

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The Role of RodA-Conserved Cysteine Residues in the Aspergillus fumigatus Conidial Surface Organization

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