Domain Shuffling between Vip3Aa and Vip3Ca: Chimera Stability and Insecticidal Activity against European, American, African, and Asian Pests
The bacterium <i>Bacillus thuringiensis</i> produces insecticidal Vip3 proteins during the vegetative growth phase with activity against several lepidopteran pests. To date, three different Vip3 protein families have been identified based on sequence identity: Vip3A, Vip3B, and Vip3C. In...
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MDPI AG
2020-02-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/12/2/99 |
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| author | Joaquín Gomis-Cebolla Rafael Ferreira dos Santos Yueqin Wang Javier Caballero Primitivo Caballero Kanglai He Juan Luis Jurat-Fuentes Juan Ferré |
| author_facet | Joaquín Gomis-Cebolla Rafael Ferreira dos Santos Yueqin Wang Javier Caballero Primitivo Caballero Kanglai He Juan Luis Jurat-Fuentes Juan Ferré |
| author_sort | Joaquín Gomis-Cebolla |
| collection | DOAJ |
| description | The bacterium <i>Bacillus thuringiensis</i> produces insecticidal Vip3 proteins during the vegetative growth phase with activity against several lepidopteran pests. To date, three different Vip3 protein families have been identified based on sequence identity: Vip3A, Vip3B, and Vip3C. In this study, we report the construction of chimeras by exchanging domains between Vip3Aa and Vip3Ca, two proteins with marked specificity differences against lepidopteran pests. We found that some domain combinations made proteins insoluble or prone to degradation by trypsin as most abundant insect gut protease. The soluble and trypsin-stable chimeras, along with the parental proteins Vip3Aa and Vip3Ca, were tested against lepidopteran pests from different continents: <i>Spodoptera exigua</i>, <i>Spodoptera littoralis</i>, <i>Spodoptera frugiperda, Helicoverpa armigera</i>, <i>Mamestra brassicae</i>, <i>Anticarsia gemmatalis</i>, and <i>Ostrinia furnacalis</i>. The exchange of the Nt domain (188 N-terminal amino acids) had little effect on the stability and toxicity (equal or slightly lower) of the resulting chimeric protein against all insects except for <i>S. frugiperda</i>, for which the chimera with the Nt domain from Vip3Aa and the rest of the protein from Vip3Ca showed a significant increase in toxicity compared to the parental Vip3Ca. Chimeras with the C-terminal domain from Vip3Aa (from amino acid 510 of Vip3Aa to the Ct) with the central domain of Vip3Ca (amino acids 189−509 based on the Vip3Aa sequence) made proteins that could not be solubilized. Finally, the chimera including the Ct domain of Vip3Ca and the Nt and central domain from Vip3Aa was unstable. Importantly, an insect species tolerant to Vip3Aa but susceptible to Vip3Ca, such as <i>Ostrinia furnacalis</i>, was also susceptible to chimeras maintaining the Ct domain from Vip3Ca, in agreement with the hypothesis that the Ct region of the protein is the one conferring specificity to Vip3 proteins. |
| first_indexed | 2024-04-11T21:43:18Z |
| format | Article |
| id | doaj.art-a638193eaff443ee941d58b57b2b852c |
| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-04-11T21:43:18Z |
| publishDate | 2020-02-01 |
| publisher | MDPI AG |
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| series | Toxins |
| spelling | doaj.art-a638193eaff443ee941d58b57b2b852c2022-12-22T04:01:32ZengMDPI AGToxins2072-66512020-02-011229910.3390/toxins12020099toxins12020099Domain Shuffling between Vip3Aa and Vip3Ca: Chimera Stability and Insecticidal Activity against European, American, African, and Asian PestsJoaquín Gomis-Cebolla0Rafael Ferreira dos Santos1Yueqin Wang2Javier Caballero3Primitivo Caballero4Kanglai He5Juan Luis Jurat-Fuentes6Juan Ferré7ERI de Biotecnología y Biomedicina (BIOTECMED), Department of Genetics, Universitat de València, 46100-Burjassot, SpainDepartment of Entomology and Plant Pathology, University of Tennessee, Knoxville, TN 37996, USAState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaInstitute for Multidisciplinary Applied Biology, Universidad Pública de Navarra, Campus Arrosadía, 31192 Mutilva, Navarra, SpainInstitute for Multidisciplinary Applied Biology, Universidad Pública de Navarra, Campus Arrosadía, 31192 Mutilva, Navarra, SpainState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaDepartment of Entomology and Plant Pathology, University of Tennessee, Knoxville, TN 37996, USAERI de Biotecnología y Biomedicina (BIOTECMED), Department of Genetics, Universitat de València, 46100-Burjassot, SpainThe bacterium <i>Bacillus thuringiensis</i> produces insecticidal Vip3 proteins during the vegetative growth phase with activity against several lepidopteran pests. To date, three different Vip3 protein families have been identified based on sequence identity: Vip3A, Vip3B, and Vip3C. In this study, we report the construction of chimeras by exchanging domains between Vip3Aa and Vip3Ca, two proteins with marked specificity differences against lepidopteran pests. We found that some domain combinations made proteins insoluble or prone to degradation by trypsin as most abundant insect gut protease. The soluble and trypsin-stable chimeras, along with the parental proteins Vip3Aa and Vip3Ca, were tested against lepidopteran pests from different continents: <i>Spodoptera exigua</i>, <i>Spodoptera littoralis</i>, <i>Spodoptera frugiperda, Helicoverpa armigera</i>, <i>Mamestra brassicae</i>, <i>Anticarsia gemmatalis</i>, and <i>Ostrinia furnacalis</i>. The exchange of the Nt domain (188 N-terminal amino acids) had little effect on the stability and toxicity (equal or slightly lower) of the resulting chimeric protein against all insects except for <i>S. frugiperda</i>, for which the chimera with the Nt domain from Vip3Aa and the rest of the protein from Vip3Ca showed a significant increase in toxicity compared to the parental Vip3Ca. Chimeras with the C-terminal domain from Vip3Aa (from amino acid 510 of Vip3Aa to the Ct) with the central domain of Vip3Ca (amino acids 189−509 based on the Vip3Aa sequence) made proteins that could not be solubilized. Finally, the chimera including the Ct domain of Vip3Ca and the Nt and central domain from Vip3Aa was unstable. Importantly, an insect species tolerant to Vip3Aa but susceptible to Vip3Ca, such as <i>Ostrinia furnacalis</i>, was also susceptible to chimeras maintaining the Ct domain from Vip3Ca, in agreement with the hypothesis that the Ct region of the protein is the one conferring specificity to Vip3 proteins.https://www.mdpi.com/2072-6651/12/2/99bacillus thuringiensisspodoptera spp., helicoverpa armigeramamestra brassicaeanticarsia gemmatalisostrinia furnacalis |
| spellingShingle | Joaquín Gomis-Cebolla Rafael Ferreira dos Santos Yueqin Wang Javier Caballero Primitivo Caballero Kanglai He Juan Luis Jurat-Fuentes Juan Ferré Domain Shuffling between Vip3Aa and Vip3Ca: Chimera Stability and Insecticidal Activity against European, American, African, and Asian Pests Toxins bacillus thuringiensis spodoptera spp., helicoverpa armigera mamestra brassicae anticarsia gemmatalis ostrinia furnacalis |
| title | Domain Shuffling between Vip3Aa and Vip3Ca: Chimera Stability and Insecticidal Activity against European, American, African, and Asian Pests |
| title_full | Domain Shuffling between Vip3Aa and Vip3Ca: Chimera Stability and Insecticidal Activity against European, American, African, and Asian Pests |
| title_fullStr | Domain Shuffling between Vip3Aa and Vip3Ca: Chimera Stability and Insecticidal Activity against European, American, African, and Asian Pests |
| title_full_unstemmed | Domain Shuffling between Vip3Aa and Vip3Ca: Chimera Stability and Insecticidal Activity against European, American, African, and Asian Pests |
| title_short | Domain Shuffling between Vip3Aa and Vip3Ca: Chimera Stability and Insecticidal Activity against European, American, African, and Asian Pests |
| title_sort | domain shuffling between vip3aa and vip3ca chimera stability and insecticidal activity against european american african and asian pests |
| topic | bacillus thuringiensis spodoptera spp., helicoverpa armigera mamestra brassicae anticarsia gemmatalis ostrinia furnacalis |
| url | https://www.mdpi.com/2072-6651/12/2/99 |
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