Domain shuffling between Vip3Aa and Vip3Ca: chimera stability and insecticidal activity against European, American, African, and Asian pests
| Autor: | Primitivo Caballero, Juan Ferré, Rafael Ferreira Dos Santos, Kanglai He, Juan Luis Jurat-Fuentes, Yueqin Wang, Javier Caballero, Joaquín Gomis-Cebolla |
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| Přispěvatelé: | Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa. IMAB - Institute for Multidisciplinary Research in Applied Biology |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Insecticides
Asia Insecta Health Toxicology and Mutagenesis medicine.medical_treatment Bacillus thuringiensis lcsh:Medicine Spodoptera Toxicology Article Lethal Dose 50 03 medical and health sciences Helicoverpa armigera Bacterial Proteins Protein Domains medicine Animals Spodoptera littoralis Pest Control Biological 030304 developmental biology chemistry.chemical_classification 0303 health sciences Protease biology 030306 microbiology Protein Stability lcsh:R fungi Spodoptera spp Ostrinia furnacalis South America biology.organism_classification Fusion protein Anticarsia gemmatalis Amino acid Europe spodoptera spp. helicoverpa armigera Biochemistry chemistry Africa North America Mamestra brassicae |
| Zdroj: | Toxins, Vol 12, Iss 2, p 99 (2020) Academica-e: Repositorio Institucional de la Universidad Pública de Navarra Universidad Pública de Navarra Toxins Volume 12 Issue 2 Academica-e. Repositorio Institucional de la Universidad Pública de Navarra instname |
| Popis: | The bacterium Bacillus thuringiensis 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: Spodoptera exigua, Spodoptera littoralis, Spodoptera frugiperda, Helicoverpa armigera, Mamestra brassicae, Anticarsia gemmatalis, and Ostrinia furnacalis. 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 S. frugiperda, 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&ndash 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 Ostrinia furnacalis, 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. |
| Databáze: | OpenAIRE |
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