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      Specific binding betweenBacillus thuringiensisCry9Aa and Vip3Aa toxins synergizes their toxicity against Asiatic rice borer (Chilo suppressalis)

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          Abstract

          <p class="first" id="d8933123e267">The bacterium <i>Bacillus thuringiensis</i> produces several insecticidal proteins, such as the crystal proteins (Cry) and the vegetative insecticidal proteins (Vip). In this work, we report that a specific interaction between two <i>B. thuringiensis</i> toxins creates insecticidal synergism and unravel the molecular basis of this interaction. When applied together, the three-domain Cry toxin Cry9Aa and the Vip Vip3Aa exhibited high insecticidal activity against an important insect pest, the Asiatic rice borer ( <i>Chilo suppressalis</i>). We found that these two proteins bind specifically to brush border membrane vesicles of <i>C. suppressalis</i> and that they do not share binding sites because no binding competition was observed between them. Binding assays revealed that the Cry9Aa and Vip3Aa proteins interacted with high affinity. We mapped their specific interacting regions by analyzing binding of Cry9Aa to overlapping fragments of Vip3Aa and by analyzing binding of Vip3Aa to individual domains of Cry9Aa. Binding to peptide arrays helped narrow the binding sites to domain II loop-3 of Cry9Aa and to <sup>428</sup>TKKMKTL <sup>434</sup> in Vip3Aa. Site-directed mutagenesis confirmed that these binding regions participate in binding that directly correlates with the synergism between the two proteins. In summary, we show that the <i>B. thuringiensis</i> Cry9Aa and Vip3Aa toxins display potent synergy based on a specific interaction between them. Our results further our understanding of the complex synergistic activities among <i>B. thuringiensis</i> toxins and are highly relevant to the development of toxin combinations for effective insect control and for delaying development of insect resistance. </p>

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          Author and article information

          Journal
          Journal of Biological Chemistry
          J. Biol. Chem.
          American Society for Biochemistry & Molecular Biology (ASBMB)
          0021-9258
          1083-351X
          July 20 2018
          July 20 2018
          July 20 2018
          June 01 2018
          : 293
          : 29
          : 11447-11458
          Article
          10.1074/jbc.RA118.003490
          6065168
          29858245
          fa34aa96-d7e1-4f2f-8fb0-3179b8954e95
          © 2018
          History

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