Infection by the fungal pathogen Cryptococcus neoformans causes lethal meningitis, primarily in immune-compromised individuals. Colonization of the brain by C. neoformans is dependent on copper (Cu) acquisition from the host, which drives critical virulence mechanisms. While C. neoformans Cu+ import and virulence are dependent on the Ctr1 and Ctr4 proteins, little is known concerning extracellular Cu ligands that participate in this process. We identified a C. neoformans gene, BIM1, that is strongly induced during Cu limitation and which encodes a protein related to lytic polysaccharide monooxygenases (LPMOs). Surprisingly, bim1 mutants are Cu deficient, and Bim1 function in Cu accumulation depends on Cu2+ coordination and cell-surface association via a glycophosphatidyl inositol anchor. Bim1 participates in Cu uptake in concert with Ctr1 and expression of this pathway drives brain colonization in mouse infection models. These studies demonstrate a role for LPMO-like proteins as a critical factor for Cu acquisition in fungal meningitis.
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The data that support the findings of this study are available from the corresponding author upon reasonable request.
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This work was partially supported by funds from the United States National Institutes of Health (NIH) (grant nos. GM041840 to D.J.T.; GM084176 to K.J.F.; GM127390 to N.V.G.), the Welch Foundation (grant no. I-1505 to N.V.G.), a postdoctoral fellowship from German Research Foundation grant PR 1727/1-1 (to C.P.), fellowship support from NIH (no. GM100678-02 to R.A.F.), NIH Molecular Mycology and Pathogenesis Training program (5T32a1052080 to A.D.S.), the Novo Nordisk Foundation grant (no. NNF17SA0027704 to K.S.J.), travel support from the School of Science and Math at the College of Charleston (to P.R.G.) and fellowship support from NIH (no. GM084146-S1) and Duke University BioCoRE (R25-GM103765) (to S.E.C.). We thank J. Lodge (Department of Molecular Microbiology, Washington University School of Medicine) for providing the anti-Cda2 antibody, Y. Song and M. Hoy for technical assistance and J.-G. Berrin for sharing information before publication. Use of the Stanford Synchrotron Radiation Light source, SLAC National Accelerator Laboratory, is supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences (contract no. DE-AC02-76SF00515). The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research, and by the NIH, National Institute of General Medical Sciences (including P41GM103393). We thank authors of works that could not be appropriately cited in this work due to space-limiting constrictions. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH.
The authors declare no competing interests.
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Garcia-Santamarina, S., Probst, C., Festa, R.A. et al. A lytic polysaccharide monooxygenase-like protein functions in fungal copper import and meningitis. Nat Chem Biol 16, 337–344 (2020). https://doi.org/10.1038/s41589-019-0437-9
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