Domain III of Cry1Ac Is Critical to Binding and Toxicity against Soybean Looper (Chrysodeixis includens) but Not to Velvetbean Caterpillar (Anticarsia gemmatalis)

Toxins
Rubina MushtaqJ L Jurat-Fuentes

Abstract

Insecticidal proteins Cry1Ac and Cry2Ac7 from the bacterium Bacillus thuringiensis (Bt) belong to the three-domain family of Bt toxins. Commercial transgenic soybean hybrids produce Cry1Ac to control the larvae of the soybean looper (Chrysodeixis includens) and the velvet bean caterpillar (Anticarsia gemmatalis). The specificity of Cry1Ac is determined by loops extending from domain II and regions of domain III in the three-dimensional structure of the toxin. In this study, we constructed a hybrid toxin (H1.2Ac) containing domains I and II of Cry1Ac and domain III of Cry2Ac7, in an attempt to obtain a protein with enhanced toxicity compared to parental toxins. Bioassays with H1.2Ac revealed toxicity against the larvae of A. gemmatalis but not against C. includens. Saturation binding assays with radiolabeled toxins and midgut brush border membrane vesicles demonstrated no specific H1.2Ac binding to C. includens, while binding in A. gemmatalis was specific and saturable. Results from competition binding assays supported the finding that Cry1Ac specificity against A. gemmatalis is mainly dictated by domain II. Taken together, these distinct interactions with binding sites may help explain the differential susceptibility to Cry1Ac ...Continue Reading

References

Jul 2, 1999·Applied and Environmental Microbiology·C RangR Laprade
Nov 9, 2000·Regulatory Toxicology and Pharmacology : RTP·F S BetzR L Fuchs
Jan 3, 2001·Applied and Environmental Microbiology·J L Jurat-Fuentes, M J Adang
Jan 5, 2002·Applied and Environmental Microbiology·Luke MassonJean-Louis Schwartz
Apr 5, 2003·Archives of Microbiology·Ruud A de MaagdSamir Naimov
Jul 22, 2004·European Journal of Biochemistry·Juan L Jurat-Fuentes, Michael J Adang
Jan 24, 2014·Bioinformatics·Philip JonesSarah Hunter

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Methods Mentioned

BETA
transgenic
saturation binding
PCR
affinity purification
Protein Assay

Software Mentioned

POLO
SigmaPlot
EMBL
PLUS
EBI InterProScan
InterProScan

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