Structural and functional role of Domain I for the insecticidal activity of the Vip3Aa protein from Bacillus thuringiensis
Abstract Vip3 proteins are produced by Bacillus thuringiensis and are toxic against lepidopterans, reason why the vip3Aa gene has been introduced into cotton and corn to control agricultural pests. Recently, the structure of Vip3 proteins has been determined and consists of a tetramer where each mon...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-10-01
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| Series: | Microbial Biotechnology |
| Online Access: | https://doi.org/10.1111/1751-7915.14110 |
| _version_ | 1798000192228687872 |
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| author | Maria Lázaro‐Berenguer Francisco Paredes‐Martínez Yolanda Bel Rafael Núñez‐Ramírez Ernesto Arias‐Palomo Patricia Casino Juan Ferré |
| author_facet | Maria Lázaro‐Berenguer Francisco Paredes‐Martínez Yolanda Bel Rafael Núñez‐Ramírez Ernesto Arias‐Palomo Patricia Casino Juan Ferré |
| author_sort | Maria Lázaro‐Berenguer |
| collection | DOAJ |
| description | Abstract Vip3 proteins are produced by Bacillus thuringiensis and are toxic against lepidopterans, reason why the vip3Aa gene has been introduced into cotton and corn to control agricultural pests. Recently, the structure of Vip3 proteins has been determined and consists of a tetramer where each monomer is composed of five structural domains. The transition from protoxin to the trypsin‐activated form involves a major conformational change of the N‐terminal Domain I, which is remodelled into a tetrameric coiled‐coil structure that is thought to insert into the apical membrane of the midgut cells. To better understand the relevance of this major change in Domain I for the insecticidal activity, we have generated several mutants aimed to alter the activity and remodelling capacity of this central region to understand its function. These mutants have been characterized by proteolytic processing, negative staining electron microscopy, and toxicity bioassays against Spodoptera exigua. The results show the crucial role of helix α1 for the insecticidal activity and in restraining the Domain I in the protoxin conformation, the importance of the remodelling of helices α2 and α3, the proteolytic processing that takes place between Domains I and II, and the role of the C‐t Domains IV and V to sustain the conformational change necessary for toxicity. |
| first_indexed | 2024-04-11T11:15:02Z |
| format | Article |
| id | doaj.art-750c753fd82b45f4b733c694992139d9 |
| institution | Directory Open Access Journal |
| issn | 1751-7915 |
| language | English |
| last_indexed | 2024-04-11T11:15:02Z |
| publishDate | 2022-10-01 |
| publisher | Wiley |
| record_format | Article |
| series | Microbial Biotechnology |
| spelling | doaj.art-750c753fd82b45f4b733c694992139d92022-12-22T04:27:14ZengWileyMicrobial Biotechnology1751-79152022-10-0115102607261810.1111/1751-7915.14110Structural and functional role of Domain I for the insecticidal activity of the Vip3Aa protein from Bacillus thuringiensisMaria Lázaro‐Berenguer0Francisco Paredes‐Martínez1Yolanda Bel2Rafael Núñez‐Ramírez3Ernesto Arias‐Palomo4Patricia Casino5Juan Ferré6Department of Genetics Universitat de València Burjassot SpainInstitut Universitari de Biotecnologia i Biomedicina BIOTECMED Universitat de València Burjassot SpainDepartment of Genetics Universitat de València Burjassot SpainCentro de Investigaciones Biológicas Margarita Salas CSIC Madrid SpainCentro de Investigaciones Biológicas Margarita Salas CSIC Madrid SpainInstitut Universitari de Biotecnologia i Biomedicina BIOTECMED Universitat de València Burjassot SpainDepartment of Genetics Universitat de València Burjassot SpainAbstract Vip3 proteins are produced by Bacillus thuringiensis and are toxic against lepidopterans, reason why the vip3Aa gene has been introduced into cotton and corn to control agricultural pests. Recently, the structure of Vip3 proteins has been determined and consists of a tetramer where each monomer is composed of five structural domains. The transition from protoxin to the trypsin‐activated form involves a major conformational change of the N‐terminal Domain I, which is remodelled into a tetrameric coiled‐coil structure that is thought to insert into the apical membrane of the midgut cells. To better understand the relevance of this major change in Domain I for the insecticidal activity, we have generated several mutants aimed to alter the activity and remodelling capacity of this central region to understand its function. These mutants have been characterized by proteolytic processing, negative staining electron microscopy, and toxicity bioassays against Spodoptera exigua. The results show the crucial role of helix α1 for the insecticidal activity and in restraining the Domain I in the protoxin conformation, the importance of the remodelling of helices α2 and α3, the proteolytic processing that takes place between Domains I and II, and the role of the C‐t Domains IV and V to sustain the conformational change necessary for toxicity.https://doi.org/10.1111/1751-7915.14110 |
| spellingShingle | Maria Lázaro‐Berenguer Francisco Paredes‐Martínez Yolanda Bel Rafael Núñez‐Ramírez Ernesto Arias‐Palomo Patricia Casino Juan Ferré Structural and functional role of Domain I for the insecticidal activity of the Vip3Aa protein from Bacillus thuringiensis Microbial Biotechnology |
| title | Structural and functional role of Domain I for the insecticidal activity of the Vip3Aa protein from Bacillus thuringiensis |
| title_full | Structural and functional role of Domain I for the insecticidal activity of the Vip3Aa protein from Bacillus thuringiensis |
| title_fullStr | Structural and functional role of Domain I for the insecticidal activity of the Vip3Aa protein from Bacillus thuringiensis |
| title_full_unstemmed | Structural and functional role of Domain I for the insecticidal activity of the Vip3Aa protein from Bacillus thuringiensis |
| title_short | Structural and functional role of Domain I for the insecticidal activity of the Vip3Aa protein from Bacillus thuringiensis |
| title_sort | structural and functional role of domain i for the insecticidal activity of the vip3aa protein from bacillus thuringiensis |
| url | https://doi.org/10.1111/1751-7915.14110 |
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