Identification, cloning and expression of a Cry1Ab cadherin receptor from European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae).
Transgenic corn expressing the Cry1Ab toxin from Bacillus thuringiensis is highly toxic to European corn borer, Ostrinia nubilalis, larvae. A putative Cry1Ab receptor (OnBt-R(1)) molecule was cloned and sequenced from a cDNA library prepared from midgut tissue of O. nubilalis larvae. The 5.6 Kb gene is homologous with a number of cadherin genes identified as Cry1 binding proteins in other lepidopterans. Brush border membrane vesicles were prepared using dissected midguts from late instars. A 220-kDa protein was identified as a cadherin-like molecule, which bound to Cry1Ab toxin and cross-reacted with an anti-cadherin serum developed from recombinant expression of a partial O. nubilalis cadherin peptide. Two additional proteins of smaller size cross-reacted with the anti-cadherin serum indicating that Cry1Ab binds to multiple receptors or to different forms of the same protein. Spodoptera frugiperda (SF9) cells transfected with the OnBt-R(1) gene were shown to express the receptor molecule which caused functional susceptibility to Cry1Ab at concentrations as low as 0.1 microg/ml. These results in combination suggest strongly that a cadherin-like protein acts as receptor and is involved with Cry1Ab toxicity in O. nubilalis.
Flannagan
Ronald D
RD
Pioneer Hi-Bred International Inc., 7301 NW 62nd Avenue, P.O. Box 85, Johnston, IA 50131-0085, USA.
Yu
Cao-Guo
CG
Mathis
John P
JP
Meyer
Terry E
TE
Shi
Xiaomei
X
Siqueira
Herbert A A
HA
Siegfried
Blair D
BD
D000595:Amino Acid Sequence; D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism*; D001427:Bacterial Toxins / Q000378:metabolism*; D015820:Cadherins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D002460:Cell Line; D004064:Digestive System / Q000737:chemistry; D004731:Endotoxins / Q000378:metabolism*; D015870:Gene Expression; D006460:Hemolysin Proteins; D019476:Insect Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D007915:Lepidoptera / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D015820:Cadherins; C095416:Cry toxin receptors; D004731:Endotoxins; D006460:Hemolysin Proteins; D019476:Insect Proteins; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis