Validation of ADAM10 metalloprotease as a Bacillus thuringiensis Cry3Aa toxin functional receptor in Colorado potato beetle (Leptinotarsa decemlineata)

Bacillus thuringiensis parasporal crystal proteins (Cry proteins) are insecticidal pore‐forming toxins that bind to specific receptor molecules on the brush border membrane of susceptible insect midgut cells to exert their toxic action. In the Colorado potato beetle (CPB), a coleopteran pest, we pre...

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Bibliographic Details
Published in:Insect molecular biology 2017-04, Vol.26 (2), p.204-214
Main Authors: Ruiz‐Arroyo, V. M., García‐Robles, I., Ochoa‐Campuzano, C., Goig, G. A., Zaitseva, E., Baaken, G., Martínez‐Ramírez, A. C., Rausell, C., Real, M. D.
Format: Article
Language:English
Subjects:
ADAM10 metalloprotease
ADAM10 Protein - metabolism
Animals
Bacillus thuringiensis
Bacterial Proteins - metabolism
Coleoptera - enzymology
Colorado potato beetle
Cry3Aa toxin
Endotoxins - metabolism
Gastrointestinal Tract - enzymology
Hemolysin Proteins - metabolism
Larva - enzymology
Leptinotarsa decemlineata
Proteolysis
Solanum tuberosum
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Summary:Bacillus thuringiensis parasporal crystal proteins (Cry proteins) are insecticidal pore‐forming toxins that bind to specific receptor molecules on the brush border membrane of susceptible insect midgut cells to exert their toxic action. In the Colorado potato beetle (CPB), a coleopteran pest, we previously proposed that interaction of Cry3Aa toxin with a CPB ADAM10 metalloprotease is an essential part of the mode of action of this toxin. Here, we annotated the gene sequence encoding an ADAM10 metalloprotease protein (CPB‐ADAM10) in the CPB genome sequencing project, and using RNA interference gene silencing we demonstrated that CPB‐ADAM10 is a Cry3Aa toxin functional receptor in CPB. Cry3Aa toxicity was significantly lower in CPB‐ADAM10 silenced larvae and in vitro toxin pore‐forming ability was greatly diminished in lipid planar bilayers fused with CPB brush border membrane vesicles (BBMVs) prepared from CPB‐ADAM10 silenced larvae. In accordance with our previous data that indicated this toxin was a substrate of ADAM10 in CPB, Cry3Aa toxin membrane‐associated proteolysis was altered when CPB BBMVs lacked ADAM10. The functional validation of CPB‐ADAM10 as a Cry3Aa toxin receptor in CPB expands the already recognized role of ADAM10 as a pathogenicity determinant of pore‐forming toxins in humans to an invertebrate species.
ISSN:0962-1075
1365-2583
DOI:10.1111/imb.12285