Processing of Candida albicans Ece1p is critical for Candidalysin maturation and fungal virulence

Jonathan P. Richardson, Selene Mogavero, David L. Moyes, Mariana Blagojevic, Thomas Kruger, Akash H. Verma, Bianca M. Coleman, Jacinto De La Cruz Diaz, Daniela Schulz, Nicole O. Ponde, Giulia Carrano, Olaf Kniemeyer, Duncan John Wilson, Oliver Bader, Simona I. Enoui, Jemima Yuer May Ho, Nessim Kichik, Sarah L Gaffen, Bernhard Hube, Julian R. Naglik

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Abstract

Candida albicans is an opportunistic fungal pathogen responsible for superficial and life threatening infections in humans. During mucosal infection, C. albicans undergoes a morphological transition from yeast to invasive filamentous hyphae that secrete Candidalysin, a 31 amino acid peptide toxin required for virulence. Candidalysin damages epithelial cell plasma membranes and stimulates the activating protein-1 (AP-1) transcription factor c-Fos (via p38- MAPK) and the MAPK phosphatase MKP1 (via ERK1/2- MAPK), which trigger and regulate pro44
inflammatory cytokine responses, respectively. The Candidalysin toxin resides as a discrete cryptic sequence within a larger 271 amino acid parental pre-pro-protein, Ece1p. Here we demonstrate that kexin-like proteinases, but not secreted aspartyl proteinases, initiate a two47 step post-translational processing of Ece1p to produce Candidalysin. Kex2p-mediated proteolysis of Ece1p after Arg61 and Arg93, but not after other processing sites within Ece1p, is required to generate immature Candidalysin from Ece1p, followed by Kex1p-mediated removal of a carboxyl arginine residue to generate mature Candidalysin. C. albicans strains harbouring mutations of Arg61 and/or Arg93 did not secrete Candidalysin, were unable to induce epithelial damage and inflammatory responses in vitro, and showed attenuated virulence in vivo in a murine model of oropharyngeal candidiasis. These observations identify enzymatic processing of C. albicans Ece1p by kexin-like proteinases as crucial steps required for Candidalysin production and fungal pathogenicity.
Original languageEnglish
Article numbere02178-17
JournalmBio
Volume9
Issue number1
DOIs
Publication statusPublished - 23 Jan 2018

Fingerprint

Candida albicans
Virulence
Activating Transcription Factor 1
Peptide Hydrolases
Dual Specificity Phosphatase 1
Cell Membrane
Aspartic Acid Proteases
Amino Acids
Hyphae
Candida albicans ECE1 protein
Candidiasis
p38 Mitogen-Activated Protein Kinases
Infection
Proteolysis
Arginine
Proteins
Yeasts
Epithelial Cells
Cytokines
Peptides

Keywords

  • candida albicans
  • candidalysin
  • fungal infection
  • kexin
  • mucosal immunity

Cite this

Richardson, J. P., Mogavero, S., Moyes, D. L., Blagojevic, M., Kruger, T., Verma, A. H., ... Naglik, J. R. (2018). Processing of Candida albicans Ece1p is critical for Candidalysin maturation and fungal virulence. mBio, 9(1), [e02178-17]. https://doi.org/10.1128/mBio.02178-17

Processing of Candida albicans Ece1p is critical for Candidalysin maturation and fungal virulence. / Richardson, Jonathan P.; Mogavero, Selene ; Moyes, David L.; Blagojevic, Mariana; Kruger, Thomas; Verma, Akash H.; Coleman, Bianca M.; Diaz, Jacinto De La Cruz; Schulz, Daniela; Ponde, Nicole O.; Carrano, Giulia; Kniemeyer, Olaf; Wilson, Duncan John; Bader, Oliver; Enoui, Simona I.; Ho, Jemima Yuer May; Kichik, Nessim; Gaffen, Sarah L; Hube, Bernhard; Naglik, Julian R.

In: mBio, Vol. 9, No. 1, e02178-17, 23.01.2018.

Research output: Contribution to journalArticle

Richardson, JP, Mogavero, S, Moyes, DL, Blagojevic, M, Kruger, T, Verma, AH, Coleman, BM, Diaz, JDLC, Schulz, D, Ponde, NO, Carrano, G, Kniemeyer, O, Wilson, DJ, Bader, O, Enoui, SI, Ho, JYM, Kichik, N, Gaffen, SL, Hube, B & Naglik, JR 2018, 'Processing of Candida albicans Ece1p is critical for Candidalysin maturation and fungal virulence', mBio, vol. 9, no. 1, e02178-17. https://doi.org/10.1128/mBio.02178-17
Richardson JP, Mogavero S, Moyes DL, Blagojevic M, Kruger T, Verma AH et al. Processing of Candida albicans Ece1p is critical for Candidalysin maturation and fungal virulence. mBio. 2018 Jan 23;9(1). e02178-17. https://doi.org/10.1128/mBio.02178-17
Richardson, Jonathan P. ; Mogavero, Selene ; Moyes, David L. ; Blagojevic, Mariana ; Kruger, Thomas ; Verma, Akash H. ; Coleman, Bianca M. ; Diaz, Jacinto De La Cruz ; Schulz, Daniela ; Ponde, Nicole O. ; Carrano, Giulia ; Kniemeyer, Olaf ; Wilson, Duncan John ; Bader, Oliver ; Enoui, Simona I. ; Ho, Jemima Yuer May ; Kichik, Nessim ; Gaffen, Sarah L ; Hube, Bernhard ; Naglik, Julian R. / Processing of Candida albicans Ece1p is critical for Candidalysin maturation and fungal virulence. In: mBio. 2018 ; Vol. 9, No. 1.
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abstract = "Candida albicans is an opportunistic fungal pathogen responsible for superficial and life threatening infections in humans. During mucosal infection, C. albicans undergoes a morphological transition from yeast to invasive filamentous hyphae that secrete Candidalysin, a 31 amino acid peptide toxin required for virulence. Candidalysin damages epithelial cell plasma membranes and stimulates the activating protein-1 (AP-1) transcription factor c-Fos (via p38- MAPK) and the MAPK phosphatase MKP1 (via ERK1/2- MAPK), which trigger and regulate pro44inflammatory cytokine responses, respectively. The Candidalysin toxin resides as a discrete cryptic sequence within a larger 271 amino acid parental pre-pro-protein, Ece1p. Here we demonstrate that kexin-like proteinases, but not secreted aspartyl proteinases, initiate a two47 step post-translational processing of Ece1p to produce Candidalysin. Kex2p-mediated proteolysis of Ece1p after Arg61 and Arg93, but not after other processing sites within Ece1p, is required to generate immature Candidalysin from Ece1p, followed by Kex1p-mediated removal of a carboxyl arginine residue to generate mature Candidalysin. C. albicans strains harbouring mutations of Arg61 and/or Arg93 did not secrete Candidalysin, were unable to induce epithelial damage and inflammatory responses in vitro, and showed attenuated virulence in vivo in a murine model of oropharyngeal candidiasis. These observations identify enzymatic processing of C. albicans Ece1p by kexin-like proteinases as crucial steps required for Candidalysin production and fungal pathogenicity.",
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author = "Richardson, {Jonathan P.} and Selene Mogavero and Moyes, {David L.} and Mariana Blagojevic and Thomas Kruger and Verma, {Akash H.} and Coleman, {Bianca M.} and Diaz, {Jacinto De La Cruz} and Daniela Schulz and Ponde, {Nicole O.} and Giulia Carrano and Olaf Kniemeyer and Wilson, {Duncan John} and Oliver Bader and Enoui, {Simona I.} and Ho, {Jemima Yuer May} and Nessim Kichik and Gaffen, {Sarah L} and Bernhard Hube and Naglik, {Julian R.}",
note = "Funding Information This work was supported by grants from the Biotechnology and Biological Sciences Research Council (BB-J016411-1, BB/N014677/1), Medical Research Council (MR/J008303/1, MR/M011372/1), FP7-PEOPLE-2013-Initial Training Network (606786), Wellcome Trust Strategic Award for Medical Mycology and Fungal Immunology (097377/Z/11/Z), and the National Institute for Health Research at Guys and St Thomas's NHS Foundation Trust and King's College London Biomedical Research Centre to JRN; the Leibniz ScienceCampus InfectoOptics SAS-2015- HKI-LWC to BH, the Deutsche Forschungsgemeinschaft (TR/CRC FungiNet, project C1 and Z2) to BH and OK, the Infect ERA-NET Program (FunComPath; BMBF 031L0001A) to BH and SM; and the National Institutes of Health (NIH; DE022550) to SLG. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Acknowledgments We thank Dr. Selvam Thavaraj for helpful discussions and Dr. Sascha Brunke for developing the “Alignator” tool that allowed us to quickly analyse LC-MS/MS output files.",
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AU - Richardson, Jonathan P.

AU - Mogavero, Selene

AU - Moyes, David L.

AU - Blagojevic, Mariana

AU - Kruger, Thomas

AU - Verma, Akash H.

AU - Coleman, Bianca M.

AU - Diaz, Jacinto De La Cruz

AU - Schulz, Daniela

AU - Ponde, Nicole O.

AU - Carrano, Giulia

AU - Kniemeyer, Olaf

AU - Wilson, Duncan John

AU - Bader, Oliver

AU - Enoui, Simona I.

AU - Ho, Jemima Yuer May

AU - Kichik, Nessim

AU - Gaffen, Sarah L

AU - Hube, Bernhard

AU - Naglik, Julian R.

N1 - Funding Information This work was supported by grants from the Biotechnology and Biological Sciences Research Council (BB-J016411-1, BB/N014677/1), Medical Research Council (MR/J008303/1, MR/M011372/1), FP7-PEOPLE-2013-Initial Training Network (606786), Wellcome Trust Strategic Award for Medical Mycology and Fungal Immunology (097377/Z/11/Z), and the National Institute for Health Research at Guys and St Thomas's NHS Foundation Trust and King's College London Biomedical Research Centre to JRN; the Leibniz ScienceCampus InfectoOptics SAS-2015- HKI-LWC to BH, the Deutsche Forschungsgemeinschaft (TR/CRC FungiNet, project C1 and Z2) to BH and OK, the Infect ERA-NET Program (FunComPath; BMBF 031L0001A) to BH and SM; and the National Institutes of Health (NIH; DE022550) to SLG. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Acknowledgments We thank Dr. Selvam Thavaraj for helpful discussions and Dr. Sascha Brunke for developing the “Alignator” tool that allowed us to quickly analyse LC-MS/MS output files.

PY - 2018/1/23

Y1 - 2018/1/23

N2 - Candida albicans is an opportunistic fungal pathogen responsible for superficial and life threatening infections in humans. During mucosal infection, C. albicans undergoes a morphological transition from yeast to invasive filamentous hyphae that secrete Candidalysin, a 31 amino acid peptide toxin required for virulence. Candidalysin damages epithelial cell plasma membranes and stimulates the activating protein-1 (AP-1) transcription factor c-Fos (via p38- MAPK) and the MAPK phosphatase MKP1 (via ERK1/2- MAPK), which trigger and regulate pro44inflammatory cytokine responses, respectively. The Candidalysin toxin resides as a discrete cryptic sequence within a larger 271 amino acid parental pre-pro-protein, Ece1p. Here we demonstrate that kexin-like proteinases, but not secreted aspartyl proteinases, initiate a two47 step post-translational processing of Ece1p to produce Candidalysin. Kex2p-mediated proteolysis of Ece1p after Arg61 and Arg93, but not after other processing sites within Ece1p, is required to generate immature Candidalysin from Ece1p, followed by Kex1p-mediated removal of a carboxyl arginine residue to generate mature Candidalysin. C. albicans strains harbouring mutations of Arg61 and/or Arg93 did not secrete Candidalysin, were unable to induce epithelial damage and inflammatory responses in vitro, and showed attenuated virulence in vivo in a murine model of oropharyngeal candidiasis. These observations identify enzymatic processing of C. albicans Ece1p by kexin-like proteinases as crucial steps required for Candidalysin production and fungal pathogenicity.

AB - Candida albicans is an opportunistic fungal pathogen responsible for superficial and life threatening infections in humans. During mucosal infection, C. albicans undergoes a morphological transition from yeast to invasive filamentous hyphae that secrete Candidalysin, a 31 amino acid peptide toxin required for virulence. Candidalysin damages epithelial cell plasma membranes and stimulates the activating protein-1 (AP-1) transcription factor c-Fos (via p38- MAPK) and the MAPK phosphatase MKP1 (via ERK1/2- MAPK), which trigger and regulate pro44inflammatory cytokine responses, respectively. The Candidalysin toxin resides as a discrete cryptic sequence within a larger 271 amino acid parental pre-pro-protein, Ece1p. Here we demonstrate that kexin-like proteinases, but not secreted aspartyl proteinases, initiate a two47 step post-translational processing of Ece1p to produce Candidalysin. Kex2p-mediated proteolysis of Ece1p after Arg61 and Arg93, but not after other processing sites within Ece1p, is required to generate immature Candidalysin from Ece1p, followed by Kex1p-mediated removal of a carboxyl arginine residue to generate mature Candidalysin. C. albicans strains harbouring mutations of Arg61 and/or Arg93 did not secrete Candidalysin, were unable to induce epithelial damage and inflammatory responses in vitro, and showed attenuated virulence in vivo in a murine model of oropharyngeal candidiasis. These observations identify enzymatic processing of C. albicans Ece1p by kexin-like proteinases as crucial steps required for Candidalysin production and fungal pathogenicity.

KW - candida albicans

KW - candidalysin

KW - fungal infection

KW - kexin

KW - mucosal immunity

U2 - 10.1128/mBio.02178-17

DO - 10.1128/mBio.02178-17

M3 - Article

VL - 9

JO - mBio

JF - mBio

SN - 2161-2129

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ER -