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Rapid, efficient auxin-inducible protein degradation in Candida pathogens

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 com...

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Autores principales: Milholland, Kedric L., Gregor, Justin B., Hoda, Smriti, Píriz-Antúnez, Soledad, Dueñas-Santero, Encarnación, Vu, Bao Gia, Patel, Krishna P., Moye-Rowley, W. Scott, Vázquez de Aldana, Carlos R., Correa-Bordes, Jaime, Briggs, Scott D., Hall, Mark C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10597344/
https://www.ncbi.nlm.nih.gov/pubmed/37594261
http://dx.doi.org/10.1128/msphere.00283-23
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author Milholland, Kedric L.
Gregor, Justin B.
Hoda, Smriti
Píriz-Antúnez, Soledad
Dueñas-Santero, Encarnación
Vu, Bao Gia
Patel, Krishna P.
Moye-Rowley, W. Scott
Vázquez de Aldana, Carlos R.
Correa-Bordes, Jaime
Briggs, Scott D.
Hall, Mark C.
author_facet Milholland, Kedric L.
Gregor, Justin B.
Hoda, Smriti
Píriz-Antúnez, Soledad
Dueñas-Santero, Encarnación
Vu, Bao Gia
Patel, Krishna P.
Moye-Rowley, W. Scott
Vázquez de Aldana, Carlos R.
Correa-Bordes, Jaime
Briggs, Scott D.
Hall, Mark C.
author_sort Milholland, Kedric L.
collection PubMed
description 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. IMPORTANCE: Life-threatening fungal infections are an escalating human health problem, complicated by limited treatment options and the evolution of drug resistant pathogen strains. Identification of new targets for therapeutics to combat invasive fungal infections, including those caused by Candida species, is an urgent need. In this report, we establish and validate an inducible protein degradation methodology in Candida albicans and Candida glabrata that provides a new tool for protein functional characterization in these, and likely other, fungal pathogen species. We expect this tool will ultimately be useful for the identification and characterization of promising drug targets and factors involved in virulence and drug resistance.
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spelling pubmed-105973442023-10-25 Rapid, efficient auxin-inducible protein degradation in Candida pathogens Milholland, Kedric L. Gregor, Justin B. Hoda, Smriti Píriz-Antúnez, Soledad Dueñas-Santero, Encarnación Vu, Bao Gia Patel, Krishna P. Moye-Rowley, W. Scott Vázquez de Aldana, Carlos R. Correa-Bordes, Jaime Briggs, Scott D. Hall, Mark C. mSphere Resource Report 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. IMPORTANCE: Life-threatening fungal infections are an escalating human health problem, complicated by limited treatment options and the evolution of drug resistant pathogen strains. Identification of new targets for therapeutics to combat invasive fungal infections, including those caused by Candida species, is an urgent need. In this report, we establish and validate an inducible protein degradation methodology in Candida albicans and Candida glabrata that provides a new tool for protein functional characterization in these, and likely other, fungal pathogen species. We expect this tool will ultimately be useful for the identification and characterization of promising drug targets and factors involved in virulence and drug resistance. American Society for Microbiology 2023-08-18 /pmc/articles/PMC10597344/ /pubmed/37594261 http://dx.doi.org/10.1128/msphere.00283-23 Text en Copyright © 2023 Milholland et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Resource Report
Milholland, Kedric L.
Gregor, Justin B.
Hoda, Smriti
Píriz-Antúnez, Soledad
Dueñas-Santero, Encarnación
Vu, Bao Gia
Patel, Krishna P.
Moye-Rowley, W. Scott
Vázquez de Aldana, Carlos R.
Correa-Bordes, Jaime
Briggs, Scott D.
Hall, Mark C.
Rapid, efficient auxin-inducible protein degradation in Candida pathogens
title Rapid, efficient auxin-inducible protein degradation in Candida pathogens
title_full Rapid, efficient auxin-inducible protein degradation in Candida pathogens
title_fullStr Rapid, efficient auxin-inducible protein degradation in Candida pathogens
title_full_unstemmed Rapid, efficient auxin-inducible protein degradation in Candida pathogens
title_short Rapid, efficient auxin-inducible protein degradation in Candida pathogens
title_sort rapid, efficient auxin-inducible protein degradation in candida pathogens
topic Resource Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10597344/
https://www.ncbi.nlm.nih.gov/pubmed/37594261
http://dx.doi.org/10.1128/msphere.00283-23
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