Transgenic Drosophila reveals a functional in vivo receptor for the Bacillus thuringiensis toxin Cry1Ac1.
The bacterium Bacillus thuringiensis synthesizes toxins (delta-endotoxins) that are highly specific for insects. Once ingested, the activated form of the toxin binds to a specific receptor(s) located on the midgut epithelial cells, inserts into the membrane causing the formation of leakage pores and eventual death of the susceptible insect larvae. Manduca sexta larvae are highly susceptible to Cry1Ac1, a toxin that is believed to bind M. sexta Aminopeptidase N, a glycoprotein located on the apical membrane. However, the binding data obtained to date only support the interaction of Cry1Ac1 with APN in vitro. To explore the in vivo role of APN, we have utilized the GAL4 enhancer trap technique to drive the expression of M. sexta APN in both midgut and mesodermal tissues of Cry1Ac1 insensitive Drosophila larvae. Transgenic Drosophila fed the toxin were now killed, demonstrating that APN can function as a receptor for Cry1Ac1 in vivo.
Gill
Michael
M
Beth Israel Deaconess Medical Center/Harvard Institutes of Medicine, Boston, Massachusetts 02115, USA. mgill2@caregroup.harvard.edu
Ellar
David
D
D000818:Animals; D030801:Animals, Genetically Modified; D001413:Bacillus thuringiensis / Q000472:pathogenicity*; D001426:Bacterial Proteins; D018076:DNA, Complementary / Q000235:genetics; D004068:Digestive System Physiological Phenomena; D004330:Drosophila / Q000382:microbiology*; D019476:Insect Proteins; D007814:Larva; D016133:Polymerase Chain Reaction; D011956:Receptors, Cell Surface / Q000235:genetics*
D001426:Bacterial Proteins; C095416:Cry toxin receptors; D018076:DNA, Complementary; D019476:Insect Proteins; D011956:Receptors, Cell Surface