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Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics

Inteins are genetic mobile elements that are inserted within protein-coding genes, which are usually housekeeping genes. They are transcribed and translated along with the host gene, then catalyze their own splicing out of the host protein, which assumes its functional conformation thereafter. As Pr...

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Autores principales: Fernandes, José Alex Lourenço, Zatti, Matheus da Silva, Arantes, Thales Domingos, de Souza, Maria Fernanda Bezerra, Santoni, Mariana Marchi, Rossi, Danuza, Zanelli, Cleslei Fernando, Liu, Xiang-Qin, Bagagli, Eduardo, Theodoro, Raquel Cordeiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9410109/
https://www.ncbi.nlm.nih.gov/pubmed/36012834
http://dx.doi.org/10.3390/jof8080846
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author Fernandes, José Alex Lourenço
Zatti, Matheus da Silva
Arantes, Thales Domingos
de Souza, Maria Fernanda Bezerra
Santoni, Mariana Marchi
Rossi, Danuza
Zanelli, Cleslei Fernando
Liu, Xiang-Qin
Bagagli, Eduardo
Theodoro, Raquel Cordeiro
author_facet Fernandes, José Alex Lourenço
Zatti, Matheus da Silva
Arantes, Thales Domingos
de Souza, Maria Fernanda Bezerra
Santoni, Mariana Marchi
Rossi, Danuza
Zanelli, Cleslei Fernando
Liu, Xiang-Qin
Bagagli, Eduardo
Theodoro, Raquel Cordeiro
author_sort Fernandes, José Alex Lourenço
collection PubMed
description Inteins are genetic mobile elements that are inserted within protein-coding genes, which are usually housekeeping genes. They are transcribed and translated along with the host gene, then catalyze their own splicing out of the host protein, which assumes its functional conformation thereafter. As Prp8 inteins are found in several important fungal pathogens and are absent in mammals, they are considered potential therapeutic targets since inhibiting their splicing would selectively block the maturation of fungal proteins. We developed a target-based drug screening system to evaluate the splicing of Prp8 intein from the yeast pathogen Cryptococcus neoformans (CnePrp8i) using Saccharomyces cerevisiae Ura3 as a non-native host protein. In our heterologous system, intein splicing preserved the full functionality of Ura3. To validate the system for drug screening, we examined cisplatin, which has been described as an intein splicing inhibitor. By using our system, new potential protein splicing inhibitors may be identified and used, in the future, as a new class of drugs for mycosis treatment. Our system also greatly facilitates the visualization of CnePrp8i splicing dynamics in vivo.
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spelling pubmed-94101092022-08-26 Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics Fernandes, José Alex Lourenço Zatti, Matheus da Silva Arantes, Thales Domingos de Souza, Maria Fernanda Bezerra Santoni, Mariana Marchi Rossi, Danuza Zanelli, Cleslei Fernando Liu, Xiang-Qin Bagagli, Eduardo Theodoro, Raquel Cordeiro J Fungi (Basel) Article Inteins are genetic mobile elements that are inserted within protein-coding genes, which are usually housekeeping genes. They are transcribed and translated along with the host gene, then catalyze their own splicing out of the host protein, which assumes its functional conformation thereafter. As Prp8 inteins are found in several important fungal pathogens and are absent in mammals, they are considered potential therapeutic targets since inhibiting their splicing would selectively block the maturation of fungal proteins. We developed a target-based drug screening system to evaluate the splicing of Prp8 intein from the yeast pathogen Cryptococcus neoformans (CnePrp8i) using Saccharomyces cerevisiae Ura3 as a non-native host protein. In our heterologous system, intein splicing preserved the full functionality of Ura3. To validate the system for drug screening, we examined cisplatin, which has been described as an intein splicing inhibitor. By using our system, new potential protein splicing inhibitors may be identified and used, in the future, as a new class of drugs for mycosis treatment. Our system also greatly facilitates the visualization of CnePrp8i splicing dynamics in vivo. MDPI 2022-08-12 /pmc/articles/PMC9410109/ /pubmed/36012834 http://dx.doi.org/10.3390/jof8080846 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Fernandes, José Alex Lourenço
Zatti, Matheus da Silva
Arantes, Thales Domingos
de Souza, Maria Fernanda Bezerra
Santoni, Mariana Marchi
Rossi, Danuza
Zanelli, Cleslei Fernando
Liu, Xiang-Qin
Bagagli, Eduardo
Theodoro, Raquel Cordeiro
Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics
title Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics
title_full Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics
title_fullStr Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics
title_full_unstemmed Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics
title_short Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics
title_sort cryptococcus neoformans prp8 intein: an in vivo target-based drug screening system in saccharomyces cerevisiae to identify protein splicing inhibitors and explore its dynamics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9410109/
https://www.ncbi.nlm.nih.gov/pubmed/36012834
http://dx.doi.org/10.3390/jof8080846
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