Rapid, efficient auxin-inducible protein degradation in

Area of Science:

• Molecular Biology
• Mycology
• Genetics

Background:

• Inducible protein degradation (IPD) systems are crucial for characterizing protein function by enabling rapid protein inactivation.
• Existing IPD tools, including the auxin-inducible degradation (AID) system, are widely used in model organisms but lack development for pathogenic fungi.
• Functional genomics in human pathogenic yeasts like Candida albicans and Candida glabrata is hindered by the absence of efficient protein manipulation tools.

Approach:

• Successfully implemented and validated both the original auxin-inducible degradation (AID) and the second-generation AID2 systems in Candida albicans and Candida glabrata.
• Developed a plasmid collection to facilitate the use of AID systems in laboratory strains of these pathogenic yeasts.
• Demonstrated rapid and efficient degradation (>95%) of target proteins within minutes using low nanomolar concentrations of 5-adamantyl-indole-3-acetic acid (5-Ad-IAA) for AID2.

Key Points:

• The AID and AID2 systems provide a rapid and efficient method for inducible protein degradation in human pathogenic yeasts.
• Auxin-induced target protein degradation effectively phenocopies gene deletion phenotypes, validating the system's utility.
• The developed systems are highly efficient, achieving >95% protein degradation rapidly and with low concentrations of the inducer.

Conclusions:

• The auxin-inducible degradation systems are now established as powerful functional genomics tools for protein characterization in Candida albicans and Candida glabrata.
• This advancement significantly enhances the study of protein function in fungal pathogens, paving the way for new therapeutic strategies.
• The adaptability of these systems suggests broad applicability to other fungal species and clinical pathogen strains.