The putative flippase Apt1 is required for intracellular membrane architecture and biosynthesis of polysaccharide and lipids in Cryptococcus neoformans

Juliana Rizzo, Ana C. Colombo, Daniel Zamith-Miranda, Vanessa K.A. Silva, Jeremy C. Allegood, Arturo Casadevall, Maurizio Del Poeta, Joshua D. Nosanchuk, James W. Kronstad, Marcio L. Rodrigues

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Flippases are responsible for the asymmetric distribution of phospholipids in biological membranes. In the encapsulated fungal pathogen Cryptococcus neoformans, the putative flippase Apt1 is an important regulator of polysaccharide secretion and pathogenesis in mice by unknown mechanisms. In this study, we analyzed the role of C. neoformans Apt1 in intracellular membrane architecture and synthesis of polysaccharide and lipids. Analysis of wild type (WT), apt1Δ (mutant) and apt1Δ::APT1 (complemented) strains by transmission electron microscopy revealed that deletion of APT1 resulted in the formation of irregular vacuoles. Disorganization of vacuolar membranes in apt1Δ cells was accompanied by a significant increase in the amounts of intra-vacuolar and pigment-containing vesicles. Quantitative immunogold labeling of C. neoformans cells with a monoclonal antibody raised to a major capsular component suggested impaired polysaccharide synthesis. APT1 deletion also affected synthesis of phosphatidylserine, phosphatidylethanolamine, inositolphosphoryl ceramide, glucosylceramide and ergosterylglycoside. These results reveal novel functions of Apt1 and are in agreement with the notion that this putative flippase plays an important role in the physiology of C. neoformans.

Original languageEnglish (US)
Pages (from-to)532-541
Number of pages10
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1865
Issue number3
DOIs
StatePublished - Mar 1 2018

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Intracellular Membranes
Cryptococcus neoformans
Polysaccharides
Lipids
Glucosylceramides
Membranes
Ceramides
Phosphatidylserines
Vacuoles
Transmission Electron Microscopy
Phospholipids
Monoclonal Antibodies

Keywords

  • Cryptococcus neoformans
  • Flippases
  • Lipid biosynthesis
  • Polysaccharide secretion
  • Vacuole

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

The putative flippase Apt1 is required for intracellular membrane architecture and biosynthesis of polysaccharide and lipids in Cryptococcus neoformans. / Rizzo, Juliana; Colombo, Ana C.; Zamith-Miranda, Daniel; Silva, Vanessa K.A.; Allegood, Jeremy C.; Casadevall, Arturo; Del Poeta, Maurizio; Nosanchuk, Joshua D.; Kronstad, James W.; Rodrigues, Marcio L.

In: Biochimica et Biophysica Acta - Molecular Cell Research, Vol. 1865, No. 3, 01.03.2018, p. 532-541.

Research output: Contribution to journalArticle

Rizzo, Juliana ; Colombo, Ana C. ; Zamith-Miranda, Daniel ; Silva, Vanessa K.A. ; Allegood, Jeremy C. ; Casadevall, Arturo ; Del Poeta, Maurizio ; Nosanchuk, Joshua D. ; Kronstad, James W. ; Rodrigues, Marcio L. / The putative flippase Apt1 is required for intracellular membrane architecture and biosynthesis of polysaccharide and lipids in Cryptococcus neoformans. In: Biochimica et Biophysica Acta - Molecular Cell Research. 2018 ; Vol. 1865, No. 3. pp. 532-541.
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