Interaction between functional domains of Bacillus thuringiensis insecticidal crystal proteins.
Interactions among the three structural domains of Bacillus thuringiensis Cry1 toxins were investigated by functional analysis of chimeric proteins. Hybrid genes were prepared by exchanging the regions coding for either domain I or domain III among Cry1Ab, Cry1Ac, Cry1C, and Cry1E. The activity of the purified trypsin-activated chimeric toxins was evaluated by testing their effects on the viability and plasma membrane permeability of Sf9 cells. Among the parental toxins, only Cry1C was active against these cells and only chimeras possessing domain II from Cry1C were functional. Combination of domain I from Cry1E with domains II and III from Cry1C, however, resulted in an inactive toxin, indicating that domain II from an active toxin is necessary, but not sufficient, for activity. Pores formed by chimeric toxins in which domain I was from Cry1Ab or Cry1Ac were slightly smaller than those formed by toxins in which domain I was from Cry1C. The properties of the pores formed by the chimeras are therefore likely to result from an interaction between domain I and domain II or III. Domain III appears to modulate the activity of the chimeric toxins: combination of domain III from Cry1Ab with domains I and II of Cry1C gave a protein which was more strongly active than Cry1C.
Rang
C
C
IGEPAM-PC, CIRAD, 34032 Montpellier Cedex 1, France.
Vachon
V
V
de Maagd
R A
RA
Villalon
M
M
Schwartz
J L
JL
Bosch
D
D
Frutos
R
R
Laprade
R
R
D000818:Animals; D001413:Bacillus thuringiensis / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D001427:Bacterial Toxins; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D018274:Electroporation; D004731:Endotoxins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D006460:Hemolysin Proteins; D002479:Inclusion Bodies; D007306:Insecticides; D018715:Microscopy, Video; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D018411:Spodoptera
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; D011993:Recombinant Fusion Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis