Phosphatidylcholine biosynthesis pathways in Cryptococcus neoformans: functional interplay and impact on virulence

As fungal diseases emerge, new studies aim to understand how different metabolic pathways, including the biosynthesis of phospholipids, influence the fungal pathogenicity. Therefore, to investigate the role of phosphatidylcholine (PC) in the biology of the human fungal pathogen Cryptococcus neoformans, a double mutant lacking OPI3 (phosphatidylethanolamine methyltransferase) from the de novo pathway, and PCT1 (choline phosphate cytidylyltransferase) from the salvage Kennedy pathway was generated using double-joint PCR coupled with biolistic technique for gene deletion. Phenotypic and virulence assays were performed, including growth viability in minimal nutrient, melanization, capsule expansion and titanization, lipid droplet analysis and in vivo infection in larval and murine models. The double mutant (opi3Δpct1Δ) exhibited normal growth in complex medium, but displayed severe growth defects and loss of viability under nutrient-limited conditions. Supplementation with L-α-glycerophosphorylcholine (GPC), PC or sorbitol fully restores growth, suggesting compensation of GPC-dependent reacylation pathway. Disruption of PC biosynthesis affected important virulence traits, including capsule formation, melanization, and titan cell development, and increased susceptibility to membrane stresses. In vivo, in both the Galleria mellonella and murine models, opi3Δpct1Δ was hypovirulent with reduced brain colonization. Other studies with C. neoformans and Candida albicans, another pathogenic yeast, showed no impact in deletion of either OPI3 or PCT1 alone for virulence and pathogenicity. Therefore, these findings highlight the critical role of PC biosynthesis for maintaining membrane integrity, morphological plasticity and host dissemination of C. neoformans.

• Publication year

  2026

• Venue

  Frontiers in Cellular and Infection Microbiology

• Publication date

  2026-01-27

• Fields of study

  Biology, Medicine, Environmental Science

• Identifiers
  DOI 10.3389/fcimb.2025.1736171PMID 41675931PMCID 12886497
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar

• No claims are published for this paper.

• No concepts are published for this paper.

• In toto imaging shows intravascular proliferation promotes transmigration of daughter cells during brain invasion of Cryptococcus neoformans.

  2025cited by this paper
• Molecular Mechanisms of Pathogenic Fungal Virulence Regulation by Cell Membrane Phospholipids

  2025cited by this paper
• Loss of Opi3 causes a lipid imbalance that influences the virulence traits of Cryptococcus neoformans but not cryptococcosis

  2024cited by this paper
• The glycerophosphocholine acyltransferase Gpc1 contributes to phosphatidylcholine biosynthesis, long-term viability, and embedded hyphal growth in Candida albicans

  2023cited by this paper
• Functional, transcriptomic, and lipidomic studies of the choC gene encoding a phospholipid methyltransferase in Aspergillus fumigatus

  2023cited by this paper
• Cryptococcus neoformans, a global threat to human health

  2023cited by this paper
• Phosphocholine cytidylyltransferase MoPct1 is crucial for vegetative growth, conidiation, and appressorium-mediated plant infection by Magnaporthe oryzae

  2023cited by this paper
• A dissemination-prone morphotype enhances extrapulmonary organ entry by Cryptococcus neoformans.

  2022cited by this paper
• Gene Expression of Diverse Cryptococcus Isolates during Infection of the Human Central Nervous System

  2021cited by this paper
• Shortening of membrane lipid acyl chains compensates for phosphatidylcholine deficiency in choline‐auxotroph yeast

  2021cited by this paper
• The Transcription Factor Pdr802 Regulates Titan Cell Formation and Pathogenicity of Cryptococcus neoformans

  2021cited by this paper
• Nutritional Conditions Modulate C. neoformans Extracellular Vesicles’ Capacity to Elicit Host Immune Response

  2020cited by this paper
• Faster Cryptococcus Melanization Increases Virulence in Experimental and Human Cryptococcosis

  2020cited by this paper
• Immunomodulatory Role of Capsular Polysaccharides Constituents of Cryptococcus neoformans

  2019cited by this paper
• Phospholipid Remodeling in Physiology and Disease.

  2019influential reference
• Phospholipid homeostasis plays an important role in fungal development, fungicide resistance and virulence in Fusarium graminearum

  2019cited by this paper
• Phosphatidylserine synthesis is essential for viability of the human fungal pathogen Cryptococcus neoformans

  2019cited by this paper
• Overproduction of Phospholipids by the Kennedy Pathway Leads to Hypervirulence in Candida albicans

  2019cited by this paper
• Variations in yeast plasma‐membrane lipid composition affect killing activity of three families of insect antifungal peptides

  2019cited by this paper
• Lipid droplet‐mediated lipid and protein homeostasis in budding yeast

  2018cited by this paper
• Maintenance of Mitochondrial Morphology in Cryptococcus neoformans Is Critical for Stress Resistance and Virulence

  2018cited by this paper
• The glycerophosphocholine acyltransferase Gpc1 is part of a phosphatidylcholine (PC)-remodeling pathway that alters PC species in yeast

  2018cited by this paper
• Lipid Biosynthesis as an Antifungal Target

  2018cited by this paper
• PCYT1A Regulates Phosphatidylcholine Homeostasis from the Inner Nuclear Membrane in Response to Membrane Stored Curvature Elastic Stress

  2018cited by this paper
• Diverse effect of phosphatidylcholine biosynthetic genes on phospholipid homeostasis, cell autophagy and fungal developments in Metarhizium robertsii

  2018cited by this paper
• From yeast to humans – roles of the Kennedy pathway for phosphatidylcholine synthesis

  2018cited by this paper
• Candida albicans Cannot Acquire Sufficient Ethanolamine from the Host To Support Virulence in the Absence of De Novo Phosphatidylethanolamine Synthesis

  2018cited by this paper
• Nile Red Staining of Neutral Lipids in Yeast.

  2017cited by this paper
• Titan cells formation in Cryptococcus neoformans is finely tuned by environmental conditions and modulated by positive and negative genetic regulators

  2017cited by this paper
• Fungal cell membrane—promising drug target for antifungal therapy

  2016cited by this paper
• Cloning of Glycerophosphocholine Acyltransferase (GPCAT) from Fungi and Plants

  2016cited by this paper
• Role for Lipid Droplet Biogenesis and Microlipophagy in Adaptation to Lipid Imbalance in Yeast.

  2015cited by this paper
• Cryptococcus neoformans and Cryptococcus gattii, the etiologic agents of cryptococcosis.

  2014cited by this paper
• Epidemiology of Cryptococcal Meningitis in the US: 1997–2009

  2013cited by this paper
• The Response to Heat Shock and Oxidative Stress in Saccharomyces cerevisiae

  2012cited by this paper
• In vitro measurement of phagocytosis and killing of Cryptococcus neoformans by macrophages.

  2012cited by this paper
• Detection of reactive oxygen species derived from the family of NOX NADPH oxidases.

  2012cited by this paper
• Titan Cells Confer Protection from Phagocytosis in Cryptococcus neoformans Infections

  2012cited by this paper
• The membrane stress response buffers lethal effects of lipid disequilibrium by reprogramming the protein homeostasis network.

  2012cited by this paper
• Phospholipid Synthesis Participates in the Regulation of Diacylglycerol Required for Membrane Trafficking at the Golgi Complex*

  2011cited by this paper
• Quantitation of cellular components in Cryptococcus neoformans for system biology analysis.

  2011cited by this paper
• The effect of L-DOPA on Cryptococcus neoformans growth and gene expression

  2011cited by this paper
• The Kennedy pathway—De novo synthesis of phosphatidylethanolamine and phosphatidylcholine

  2010cited by this paper
• Regulation of phospholipid synthesis in yeast This work was supported in part by the United States Public Health Service, National Institutes of Health Grants GM-28140 and GM-50679. Published, JLR Papers in Press, October 27, 2008.

  2009cited by this paper
• Vesicle-associated melanization in Cryptococcus neoformans.

  2009cited by this paper
• Depletion of phosphatidylcholine affects endoplasmic reticulum morphology and protein traffic at the Golgi complex

  2009cited by this paper
• An efficient gene-disruption method in Cryptococcus neoformans by double-joint PCR with NAT-split markers.

  2009cited by this paper
• Membrane lipids: where they are and how they behave

  2008cited by this paper
• Extracellular Vesicles Produced by Cryptococcus neoformans Contain Protein Components Associated with Virulence

  2007cited by this paper
• Identification of fungal cell wall mutants using susceptibility assays based on Calcofluor white and Congo red

  2006cited by this paper
• Genomic Analysis of the Opi− Phenotype

  2006cited by this paper
• Cryptococcus neoformans α Strains Preferentially Disseminate to the Central Nervous System during Coinfection

  2005cited by this paper
• Depletion of phosphatidylcholine in yeast induces shortening and increased saturation of the lipid acyl chains: evidence for regulation of intrinsic membrane curvature in a eukaryote.

  2005cited by this paper
• Regulatory enzymes of phosphatidylcholine biosynthesis: a personal perspective.

  2005influential reference
• Galleria mellonella as a Model System To Study Cryptococcus neoformans Pathogenesis

  2005cited by this paper
• Novel gene functions required for melanization of the human pathogen Cryptococcus neoformans

  2005cited by this paper
• Regulation of Phospholipid Synthesis in the Yeastcki1Δeki1Δ Mutant Defective in the Kennedy Pathway: THECHO1-ENCODED PHOSPHATIDYLSERINE SYNTHASE IS REGULATED BY mRNA STABILITY

  2004cited by this paper
• Regulation of phospholipid synthesis in the yeast cki1Delta eki1Delta mutant defective in the Kennedy pathway. The Cho1-encoded phosphatidylserine synthase is regulated by mRNA stability.

  2004cited by this paper
• Recapitulation of the Sexual Cycle of the Primary Fungal Pathogen Cryptococcus neoformans var. gattii: Implications for an Outbreak on Vancouver Island, Canada

  2003cited by this paper
• Identification of metabolites of importance in the pathogenesis of pulmonary cryptococcoma using nuclear magnetic resonance spectroscopy.

  2003cited by this paper
• Regulation of vesicle trafficking, transcription, and meiosis: lessons learned from yeast regarding the disparate biologies of phosphatidylcholine.

  2001cited by this paper
• Cryptococcomas distinguished from gliomas with MR spectroscopy: an experimental rat and cell culture study.

  2001influential reference
• The Yeast Cell Wall and Septum as Paradigms of Cell Growth and Morphogenesis*

  2001cited by this paper
• Phospholipid biosynthesis in the yeast Saccharomyces cerevisiae and interrelationship with other metabolic processes.

  1999cited by this paper
• Phosphatidylinositol signalling reactions.

  1998cited by this paper
• Mitochondrial function is required for resistance to oxidative stress in the yeast Saccharomyces cerevisiae

  1997cited by this paper
• Susceptibility of melanized and nonmelanized Cryptococcus neoformans to nitrogen- and oxygen-derived oxidants

  1994cited by this paper
• Separation and analysis of phospholipids by thin layer chromatography.

  1993cited by this paper
• Lipids in the Structure and Function of Fungal Membranes

  1990cited by this paper

• No citing papers are available for this paper.