The receptor of Bacillus sphaericus binary toxin in Culex pipiens (Diptera: Culicidae) midgut: molecular cloning and expression.
Culex pipiens larval midgut is the primary target of the binary toxin (Bin) present in parasporal inclusions of Bacillus sphaericus. Cpm1, a 60-kDa protein purified from brush border membranes, has been proposed as the receptor of the Bin toxin in the midgut epithelial cells of mosquitoes. We have cloned and characterized the corresponding cDNA from midgut of Culex pipiens larvae. The open reading frame predicted a 580 amino-acid protein with a putative signal peptide at the N-terminus and a putative GPI-anchoring signal at the C-terminus. The amino acid sequence of the cloned Cpm1 exhibited 39-43% identities with insect maltases (alpha-glucosidases and alpha-amylases). Recombinant Cpm1 expressed in E. coli specifically bound to the Bin toxin and had a significant alpha-glucosidase activity but no alpha-amylase activity. These results support the view that Cpm1 is an alpha-glucosidase expressed in Culex midgut where it constitutes the receptor for the Bin toxin. To date, this is the first component involved in the mosquitocidal activity of the Bacillus sphaericus Bin toxin to be characterized. Its identification provides a key step to elucidate the mode of action of the Bin toxin and the mechanisms of resistance developed against it by some mosquito strains.
Darboux
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I
Institut National de la Recherche Agronomique, Unité Santé Végétale et Environnement, 123, Boulevard Francis Meilland, 06606 Antibes Cedex, France.
Nielsen-LeRoux
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C
Charles
J F
JF
Pauron
D
D
D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000378:metabolism*; D001483:Base Sequence; D015152:Blotting, Northern / Q000379:methods; D003001:Cloning, Molecular; D003465:Culex / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary; D004064:Digestive System; D015870:Gene Expression; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D000520:alpha-Glucosidases
C405322:Bacillus sphaericus binary toxin receptor; D018076:DNA, Complementary; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D000520:alpha-Glucosidases