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Related Experiment Video

Updated: Jun 9, 2025

Use of In Vivo Imaging to Screen for Morphogenesis Phenotypes in Candida albicans Mutant Strains During Active Infection in a Mammalian Host

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Alternative isoforms and phase separation of Ref1 repress morphogenesis in Cryptococcus.

Nathan K Glueck¹, Xiaofeng Xie¹, Xiaorong Lin¹

¹Department of Microbiology, University of Georgia, Athens, GA, USA.

|October 30, 2024

Summary

Researchers identified Ref1, a repressor that completes a negative feedback loop regulating fungal development in Cryptococcus neoformans. This mechanism uses isoform-specific phase separation to control the yeast-to-hypha transition.

Keywords:

CP: Cell biology CP: Microbiology Cryptococcus neoformans alternative transcription feedback regulation morphogenesis phase separation sexual development

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Last Updated: Jun 9, 2025

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Area of Science:

Fungal Biology
Molecular Biology
Cellular Biology

Background:

Cryptococcus neoformans serves as a model organism for eukaryotic and fungal biology.
Negative feedback loops are crucial for regulating developmental transitions in eukaryotes.
Understanding these regulatory mechanisms is key to deciphering fungal morphogenesis.

Purpose of the Study:

To identify and characterize novel regulators of the yeast-to-hypha transition in Cryptococcus neoformans.
To elucidate the role of negative feedback in fungal sexual development.
To investigate the functional implications of alternative transcription and protein isoform diversity.

Main Methods:

Investigated the yeast-to-hypha transition in Cryptococcus neoformans.
Identified and characterized the repressor protein Ref1.
Analyzed the role of the master regulator Znf2 in regulating Ref1 transcription.
Examined the impact of alternative transcription on Ref1 isoform function.
Studied the influence of phase separation on Ref1 activity and degradation.

Main Results:

Discovered Ref1 as a repressor of the yeast-to-hypha transition, completing a negative feedback loop initiated by Znf2.
Demonstrated that alternative transcription of Ref1, driven by Znf2, produces functionally distinct isoforms.
Showed that isoform-specific phase separation capacity influences repressor strength and proteolytic susceptibility.
Confirmed that Ref1's multimodal nature allows for context-dependent activity tuning.

Conclusions:

Ref1 completes a negative feedback loop regulating hyphal morphogenesis during sexual development in Cryptococcus neoformans.
Alternative transcription and isoform-specific phase separation provide a mechanism for fine-tuning gene repression.
This study reveals how phase separation enables a transcriptional program to modulate its own repression, guiding morphological transitions.