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Functional characterizations of residues Arg-158 and Tyr-170 of the mosquito-larvicidal Bacillus thuringiensis Cry4Ba

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초록

The insecticidal activity of Bacillus thuringiensis (Bt) Cry toxinsinvolves toxin stabilization, oligomerization, passage acrossthe peritrophic membrane (PM), binding to midgut receptorsand pore-formation. The residues Arg-158 and Tyr-170 havebeen shown to be crucial for the toxicity of Bt Cry4Ba. Wecharacterized the biological function of these residues. Inmosquito larvae, the mutants R158A/E/Q (R158) could hardlypenetrate the PM due to a significantly reduced ability to alterPM permeability; the mutant Y170A, however, could passthrough the PM, but degraded in the space between the PMand the midgut epithelium. Further characterization byoligomerization demonstrated that Arg-158 mutants failed toform correctly sized high-molecular weight oligomers. This isthe first report that Arg-158 plays a role in the formation ofCry4Ba oligomers, which are essential for toxin passage acrossthe PM. Tyr-170, meanwhile, is involved in toxin stabilization in the toxic mechanism of Cry4Ba in mosquito larvae.

Abstract

The insecticidal activity of Bacillus thuringiensis (Bt) Cry toxinsinvolves toxin stabilization, oligomerization, passage acrossthe peritrophic membrane (PM), binding to midgut receptorsand pore-formation. The residues Arg-158 and Tyr-170 havebeen shown to be crucial for the toxicity of Bt Cry4Ba. Wecharacterized the biological function of these residues. Inmosquito larvae, the mutants R158A/E/Q (R158) could hardlypenetrate the PM due to a significantly reduced ability to alterPM permeability; the mutant Y170A, however, could passthrough the PM, but degraded in the space between the PMand the midgut epithelium. Further characterization byoligomerization demonstrated that Arg-158 mutants failed toform correctly sized high-molecular weight oligomers. This isthe first report that Arg-158 plays a role in the formation ofCry4Ba oligomers, which are essential for toxin passage acrossthe PM. Tyr-170, meanwhile, is involved in toxin stabilization in the toxic mechanism of Cry4Ba in mosquito larvae.

참고문헌 (27)

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