P-type atpase apt1 of the fungal pathogen Cryptococcus neoformans is a lipid flippase of broad substrate specificity
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- P-Type ATPase Apt1 of the Fungal Pathogen Cryptococcus neoformans Is a Lipid Flippase of Broad Substrate Specificity
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Lipid flippases of the P4-ATPase family are ATP-driven transporters that translocate lipids from the exoplasmic to the cytosolic leaflet of biological membranes. In the encapsulated fungal pathogen Cryptococcus neoformans, the P4-ATPase Apt1p is an important regulator of polysaccharide secretion and pathogenesis, but its biochemical characterization is lacking. Phylogenetic analysis revealed that Apt1p belongs to the subclade of P4A-ATPases characterized by the common requirement for a β-subunit. Using heterologous expression in S. cerevisiae, we demonstrate that Apt1p forms a heterodimeric complex with the C. neoformans Cdc50 protein. This association is required for both localization and activity of the transporter complex. Lipid flippase activity of the heterodimeric complex was assessed by complementation tests and uptake assays employing fluorescent lipids and revealed a broad substrate specificity, including several phospholipids, the alkylphospholipid miltefosine, and the glycolipids glucosyl-and galactosylceramide. Our results suggest that transbilayer lipid transport in C. neoformans is finely regulated to promote fungal virulence, which reinforces the potential of Apt1p as a target for antifungal drug development.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 843 |
| Tidsskrift | Journal of Fungi |
| Vol/bind | 7 |
| Udgave nummer | 10 |
| Antal sider | 16 |
| DOI | |
| Status | Udgivet - 2021 |
Bibliografisk note
Funding Information:
Funding: This work was funded by grants from the Novo Nordisk Fonden (NNF18OC0034784) and the German Research Foundation (INST 213/886-1 FUGG) to T.G.P. and L.D.S.; R.P. and L.A.P. acknowledge funding from the Research Exchange Program of the Ruhr University Bochum. J.R. acknowledges funding from the Research Explorer Ruhr (Research Exchange Program of the Ruhr University Bochum) conducted by Research School PLUS, Germany’s Excellence Initiative [DFG GSC 98/3] and the Pasteur-Roux-Cantarini fellowship of Institute Pasteur. S.V. is a scholar of the Studienstiftung des Deutschen Volkes.
Funding Information:
This work was funded by grants from the Novo Nordisk Fonden (NNF18OC0034784) and the German Research Foundation (INST 213/886-1 FUGG) to T.G.P. and L.D.S.; R.P. and L.A.P. acknowledge funding from the Research Exchange Program of the Ruhr University Bochum. J.R. acknowledges funding from the Research Explorer Ruhr (Research Exchange Program of the Ruhr University Bochum) conducted by Research School PLUS, Germany?s Excellence Initiative [DFG GSC 98/3] and the Pasteur-Roux-Cantarini fellowship of Institute Pasteur. S.V. is a scholar of the Studienstiftung des Deutschen Volkes. Acknowledgments: The authors are grateful to James Kronstad for sharing ideas in early studies on the role of lipid flippases in Cryptococcus. Imaging was in part carried out at the Center for Advanced Bioimaging Denmark (CAB).
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
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