<i>Cryptococcus neoformans</i> Prp8 Intein: An In Vivo Target-Based Drug Screening System in <i>Saccharomyces cerevisiae</i> 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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| Format: | Article |
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MDPI AG
2022-08-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/8/8/846 |
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| author | José Alex Lourenço Fernandes Matheus da Silva Zatti Thales Domingos Arantes Maria Fernanda Bezerra de Souza Mariana Marchi Santoni Danuza Rossi Cleslei Fernando Zanelli Xiang-Qin Liu Eduardo Bagagli Raquel Cordeiro Theodoro |
| author_facet | José Alex Lourenço Fernandes Matheus da Silva Zatti Thales Domingos Arantes Maria Fernanda Bezerra de Souza Mariana Marchi Santoni Danuza Rossi Cleslei Fernando Zanelli Xiang-Qin Liu Eduardo Bagagli Raquel Cordeiro Theodoro |
| author_sort | José Alex Lourenço Fernandes |
| collection | DOAJ |
| 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 <i>Cryptococcus neoformans</i> (CnePrp8i) using <i>Saccharomyces cerevisiae</i> 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. |
| first_indexed | 2024-03-09T13:08:28Z |
| format | Article |
| id | doaj.art-39aa3f9e192d4ec4a644d451fca174e1 |
| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-09T13:08:28Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-39aa3f9e192d4ec4a644d451fca174e12023-11-30T21:45:07ZengMDPI AGJournal of Fungi2309-608X2022-08-018884610.3390/jof8080846<i>Cryptococcus neoformans</i> Prp8 Intein: An In Vivo Target-Based Drug Screening System in <i>Saccharomyces cerevisiae</i> to Identify Protein Splicing Inhibitors and Explore Its DynamicsJosé Alex Lourenço Fernandes0Matheus da Silva Zatti1Thales Domingos Arantes2Maria Fernanda Bezerra de Souza3Mariana Marchi Santoni4Danuza Rossi5Cleslei Fernando Zanelli6Xiang-Qin Liu7Eduardo Bagagli8Raquel Cordeiro Theodoro9Institute of Tropical Medicine, Federal University of Rio Grande do Norte (UFRN), Natal 59077-080, Rio Grande do Norte, BrazilInstitute of Tropical Medicine, Federal University of Rio Grande do Norte (UFRN), Natal 59077-080, Rio Grande do Norte, BrazilDepartment of Biosciences and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goiás, Goiânia 74605-050, Goiás, BrazilDepartment of Biochemistry, Biosciences Center, Federal University of Rio Grande do Norte (UFRN), Natal 59078-900, Rio Grande do Norte, BrazilSchool of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara 14800-903, São Paulo, BrazilPensabio, São Paulo 05005-010, São Paulo, BrazilSchool of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara 14800-903, São Paulo, BrazilDepartment of Biochemistry and Molecular Biology, Dalhousie University, Halifax, NS B3H 4R2, CanadaMicrobiology and Immunology Department, Biosciences Institute of Botucatu, São Paulo State University (UNESP), Botucatu 18618-689, São Paulo, BrazilInstitute of Tropical Medicine, Federal University of Rio Grande do Norte (UFRN), Natal 59077-080, Rio Grande do Norte, BrazilInteins 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 <i>Cryptococcus neoformans</i> (CnePrp8i) using <i>Saccharomyces cerevisiae</i> 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.https://www.mdpi.com/2309-608X/8/8/846protein splicinginteinPrp8Ura3CnePrp8icisplatin |
| spellingShingle | José Alex Lourenço Fernandes Matheus da Silva Zatti Thales Domingos Arantes Maria Fernanda Bezerra de Souza Mariana Marchi Santoni Danuza Rossi Cleslei Fernando Zanelli Xiang-Qin Liu Eduardo Bagagli Raquel Cordeiro Theodoro <i>Cryptococcus neoformans</i> Prp8 Intein: An In Vivo Target-Based Drug Screening System in <i>Saccharomyces cerevisiae</i> to Identify Protein Splicing Inhibitors and Explore Its Dynamics Journal of Fungi protein splicing intein Prp8 Ura3 CnePrp8i cisplatin |
| title | <i>Cryptococcus neoformans</i> Prp8 Intein: An In Vivo Target-Based Drug Screening System in <i>Saccharomyces cerevisiae</i> to Identify Protein Splicing Inhibitors and Explore Its Dynamics |
| title_full | <i>Cryptococcus neoformans</i> Prp8 Intein: An In Vivo Target-Based Drug Screening System in <i>Saccharomyces cerevisiae</i> to Identify Protein Splicing Inhibitors and Explore Its Dynamics |
| title_fullStr | <i>Cryptococcus neoformans</i> Prp8 Intein: An In Vivo Target-Based Drug Screening System in <i>Saccharomyces cerevisiae</i> to Identify Protein Splicing Inhibitors and Explore Its Dynamics |
| title_full_unstemmed | <i>Cryptococcus neoformans</i> Prp8 Intein: An In Vivo Target-Based Drug Screening System in <i>Saccharomyces cerevisiae</i> to Identify Protein Splicing Inhibitors and Explore Its Dynamics |
| title_short | <i>Cryptococcus neoformans</i> Prp8 Intein: An In Vivo Target-Based Drug Screening System in <i>Saccharomyces cerevisiae</i> to Identify Protein Splicing Inhibitors and Explore Its Dynamics |
| title_sort | i cryptococcus neoformans i prp8 intein an in vivo target based drug screening system in i saccharomyces cerevisiae i to identify protein splicing inhibitors and explore its dynamics |
| topic | protein splicing intein Prp8 Ura3 CnePrp8i cisplatin |
| url | https://www.mdpi.com/2309-608X/8/8/846 |
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