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:10.1101/2023.05.17.541235

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Rapid, efficient auxin-inducible protein degradation in Candida pathogens

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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, …

bioRxiv

Cold Spring Harbor Laboratory

05/18/2023

DOI: 10.1101/2023.05.17.541235

PMCID: PMC10245712

PMID: 37293017

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https://doi.org/10.1101/2023.05.17.541235View

Preprint (Author's original)This preprint has not been evaluated by subject experts through peer review. Preprints may undergo extensive changes and/or become peer-reviewed journal articles. Open Access

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 (5-Ad-IAA). 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 - Purdue University System
Justin B. Gregor - Purdue University System
Smriti Hoda - Purdue University System
Soledad Píriz-Antúnez - Universidad de Extremadura
Encarnación Dueñas-Santero - Universidad de Salamanca
Bao Gia Vu - University of Iowa
Krishna P. Patel - Purdue University System
W. Scott Moye-Rowley - University of Iowa
Carlos R. Vázquez de Aldana
Jaime Correa-Bordes - Universidad de Extremadura
Scott D. Briggs - Purdue University System
Mark C. Hall - Purdue University System
Resource Type: Preprint
Publication Details: bioRxiv
DOI: 10.1101/2023.05.17.541235
PMID: 37293017
PMCID: PMC10245712
Publisher: Cold Spring Harbor Laboratory
Language: English
Date posted: 05/18/2023
Academic Unit: Molecular Physiology and Biophysics
Record Identifier: 9984426852402771

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