title: Validation of ADAM10 metalloprotease as a Bacillus thuringiensis Cry3Aa toxin functional receptor in Colorado potato beetle (Leptinotarsa decemlineata).
abstract: 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.
authors.affiliation: Department of Genetics, University of Valencia, Burjassot, Valencia, Spain.
authors.forename: V M
authors.initials: VM
authors.lastname: Ruiz-Arroyo
authors.affiliation: Department of Genetics, University of Valencia, Burjassot, Valencia, Spain.
authors.forename: I
authors.identifier: 0000-0002-0165-399X
authors.initials: I
authors.lastname: García-Robles
authors.affiliation: Department of Genetics, University of Valencia, Burjassot, Valencia, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Ochoa-Campuzano
authors.affiliation: Department of Genetics, University of Valencia, Burjassot, Valencia, Spain.
authors.forename: G A
authors.initials: GA
authors.lastname: Goig
authors.affiliation: Department of Physiology, University of Freiburg, Freiburg, Germany.|Ionera Technologies GmbH, Freiburg, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Zaitseva
authors.affiliation: Ionera Technologies GmbH, Freiburg, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Baaken
authors.affiliation: Servicios Centrales de Soporte a la Investigación Experimental (SCSIE), University of Valencia, Burjassot, Valencia, Spain.
authors.forename: A C
authors.initials: AC
authors.lastname: Martínez-Ramírez
authors.affiliation: Department of Genetics, University of Valencia, Burjassot, Valencia, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Rausell
authors.affiliation: Department of Genetics, University of Valencia, Burjassot, Valencia, Spain.
authors.forename: M D
authors.initials: MD
authors.lastname: Real
mesh_terms: D000072197:ADAM10 Protein / Q000378:metabolism*; D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism*; D001517:Coleoptera / Q000201:enzymology*; D004731:Endotoxins / Q000378:metabolism*; D041981:Gastrointestinal Tract / Q000201:enzymology; D006460:Hemolysin Proteins / Q000378:metabolism*; D007814:Larva / Q000201:enzymology; D059748:Proteolysis
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D000072197:ADAM10 Protein
keywords: ADAM10 metalloprotease; Bacillus thuringiensis; Colorado potato beetle; Cry3Aa toxin