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Journal of Bacteriology, November 2002, p. 5848-5854, Vol. 184, No. 21
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.21.5848-5854.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

An ABC Transporter from Bacillus thuringiensis Is Essential for ß-Exotoxin I Production

Sylvain Espinasse,1 Michel Gohar,1,2 Didier Lereclus,1,3 and Vincent Sanchis1,3*

Unité de Recherches de Lutte Biologique, INRA La Minière, 78285 Guyancourt,1 Unité de Biochimie Microbienne, Institut Pasteur, 75724 Paris Cedex 15, France,3 Aventis Crop Science, B 9000 Ghent, Belgium2

Received 12 April 2002/ Accepted 6 August 2002

ß-Exotoxin I is a nonspecific insecticidal metabolite secreted by some Bacillus thuringiensis strains. Several studies of B. thuringiensis strains that have lost the capacity to produce ß-exotoxin I have suggested that there is a strong correlation between high levels of ß-exotoxin I production and the ability to synthesize crystal proteins. In this study, we showed that a mutant strain, B. thuringiensis 407-1(Cry-)(Pig+), with no crystal gene, produced considerable amounts of ß-exotoxin I together with a soluble brown melanin pigment. Therefore, ß-exotoxin I production can take place after a strain has lost the plasmids bearing the cry genes, which suggests that these curable plasmids probably contain determinants involved in the regulation of ß-exotoxin I production. Using a mini-Tn10 transposon, we constructed a library of strain 407-1(Cry-)(Pig+) mutants. We screened for nonpigmented mutants with impaired ß-exotoxin I production and identified a genetic locus harboring two genes (berA and berB) essential for ß-exotoxin I production. The deduced amino acid sequence of the berA gene displayed significant similarity to the ATP-binding domains of the DRI (drug resistance and immunity) family of ATP-binding cassette (ABC) proteins involved in drug resistance and immunity to bacteriocins and lantibiotics. The berB gene encodes a protein with six putative transmembrane helices, which probably constitutes the integral membrane component of the transporter. The demonstration that berAB is required for ß-exotoxin I production and/or resistance in B. thuringiensis adds an adenine nucleotide analog to the wide range of substrates of the superfamily of ABC proteins. We suggest that berAB confers ß-exotoxin I immunity in B. thuringiensis, through active efflux of the molecule.


* Corresponding author. Mailing address: Unité de Biochimie Microbienne, Institut Pasteur, 25/28 rue du Dr. Roux, 75724 Paris Cedex 15, France. Phone: 33 1 45 68 88 12. Fax: 33 1 45 68 89 38. E-mail: vsanchis{at}pasteur.fr.


Journal of Bacteriology, November 2002, p. 5848-5854, Vol. 184, No. 21
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.21.5848-5854.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.




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