Fluorescent organelle markers in Cryptococcus neoformans: a versatile toolkit for live-cell subcellular localization

Area of Science:

• Mycology
• Cell Biology
• Molecular Biology

Background:

• Intracellular organelle dynamics are crucial for fungal growth, differentiation, and pathogenicity.
• The fungal pathogen Cryptococcus neoformans shows diverse morphology, but organelle roles in adaptation are unclear.

Purpose of the Study:

• To create and validate a toolkit for high-resolution live-cell imaging of ten subcellular compartments in Cryptococcus neoformans.
• To investigate organelle remodeling during fungal adaptation to host-relevant conditions and sexual development.

Main Methods:

• Developed fluorescent organelle marker plasmids and strains (mCherry/GFP fusions).
• Utilized high-resolution live-cell imaging for ten major organelles.
• Performed colocalization analyses using diploid formation and dual-labeling.

Main Results:

• Established a comprehensive imaging platform for Cryptococcus neoformans organelles.
• Revealed condition- and stage-specific organelle architecture remodeling during vegetative growth and mating.
• Demonstrated the toolkit's utility for analyzing organelle dynamics under various conditions.

Conclusions:

• The developed imaging platform provides a robust framework for studying fungal adaptation, development, and virulence.
• Organelle organization is dynamically remodeled, supporting fungal adaptation and pathogenicity.
• The toolkit facilitates functional inference of uncharacterized genes based on spatial localization.