Rapid, efficient auxin-inducible protein degradation in Candida pathogens

Kedric L Milholland; Justin B Gregor; Smriti Hoda; Soledad Píriz-Antúnez; Encarnación Dueñas-Santero; Bao Gia Vu; Krishna P Patel; W Scott Moye-Rowley; Carlos R Vázquez de Aldana; Jaime Correa-Bordes; Scott D Briggs; Mark C Hall

doi:10.1128/msphere.00283-23

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Rapid, efficient auxin-inducible protein degradation in Candida pathogens

Journal article

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Rapid, efficient auxin-inducible protein degradation in Candida pathogens

Kedric L Milholland, Justin B Gregor, Smriti Hoda, Soledad Píriz-Antúnez, Encarnación Dueñas-Santero, Bao Gia Vu, Krishna P Patel, W Scott Moye-Rowley, Carlos R Vázquez de Aldana, Jaime Correa-Bordes, …

mSphere, Vol.8(5), e00283-23

10/24/2023

DOI: 10.1128/msphere.00283-23

PMCID: PMC10597344

PMID: 37594261

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https://doi.org/10.1128/msphere.00283-23View

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Abstract

A variety of inducible protein degradation (IPD) systems have been developed as powerful tools for protein functional characterization. IPD systems provide a convenient mechanism for rapid inactivation of almost any target protein of interest. Auxin-inducible degradation (AID) is one of the most common IPD systems and has been established in diverse eukaryotic research model organisms. Thus far, IPD tools have not been developed for use in pathogenic fungal species. Here, we demonstrate that the original AID and the second generation, AID2, systems work efficiently and rapidly in the human pathogenic yeasts, Candida albicans and Candida glabrata. We developed a collection of plasmids that support AID system use in laboratory strains of these pathogens. These systems can induce >95% degradation of target proteins within minutes. In the case of AID2, maximal degradation was achieved at low nanomolar concentrations of the synthetic auxin analog 5-adamantyl-indole-3-acetic acid. Auxin-induced target degradation successfully phenocopied gene deletions in both species. The system should be readily adaptable to other fungal species and to clinical pathogen strains. Our results define the AID system as a powerful and convenient functional genomics tool for protein characterization in fungal pathogens.

Details

Title: Subtitle: Rapid, efficient auxin-inducible protein degradation in Candida pathogens
Creators: Kedric L Milholland
Justin B Gregor
Smriti Hoda
Soledad Píriz-Antúnez
Encarnación Dueñas-Santero
Bao Gia Vu
Krishna P Patel
W Scott Moye-Rowley
Carlos R Vázquez de Aldana
Jaime Correa-Bordes
Scott D Briggs
Mark C Hall
Resource Type: Journal article
Publication Details: mSphere, Vol.8(5), e00283-23
DOI: 10.1128/msphere.00283-23
PMID: 37594261
PMCID: PMC10597344
NLM abbreviation: mSphere
ISSN: 2379-5042
eISSN: 2379-5042
Grant note: DOI: 10.13039/100000060, name: HHS | NIH | National Institute of Allergy and Infectious Diseases, award: AI136995; DOI: 10.13039/100000060, name: HHS | NIH | National Institute of Allergy and Infectious Diseases, award: AI152494; DOI: 10.13039/100000060, name: HHS | NIH | National Institute of Allergy and Infectious Diseases, award: AI168050; DOI: 10.13039/100000060, name: HHS | NIH | National Institute of Allergy and Infectious Diseases, award: T32AI148103; DOI: 10.13039/100006975, name: Indiana Clinical and Translational Sciences Institute, award: UL1TR002529; name: MEC | Agencia Estatal de Investigación, award: PID2020-118109RB-I00; name: EC | European Regional Development Fund, award: CL-EI-2021-08-IBFG
Language: English
Electronic publication date: 08/18/2023
Date published: 10/24/2023
Academic Unit: Molecular Physiology and Biophysics
Record Identifier: 9984455946602771

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