A Novel Secreted Protein Toxin from the Insect Pathogenic Bacterium Xenorhabdus nematophila.pdf

A Novel Secreted Protein Toxin from the Insect Pathogenic Bacterium Xenorhabdus nematophila* Received for publication, September 5, 2003, and in revised form, December 19, 2003 Published, JBC Papers in Press, January 5, 2004, DOI 10.1074/jbc.M309859200 Susan E. Brown?, Anh T. Cao, Eric R. Hines, Raymond J. Akhurst, and Peter D. East From the CSIRO Entomology, GPO Box 1700, Acton, Australian Capital Territory 2601, Australia The bacterium Xenorhabdus nematophila is an insect pathogen that produces several proteins that enable it to kill insects. Screening of a cosmid library constructed from X. nematophila strain A24 identified a gene that encoded a novel protein that was toxic to insects. The 42-kDa protein encoded by the toxin gene was expressed and purified from a recombinant system, and was shown to kill the larvae of insects such as Galleria mellonella and Helicoverpa armigera when injected at doses of around 30–40 ng/g larvae. Sequencing and bioinfor- matic analysis suggested that the toxin was a novel pro- tein, and that it was likely to be part of a genomic island involved in pathogenicity. When the native bacteria were grown under laboratory conditions, a soluble form of the 42-kDa toxin was secreted only by bacteria in the phase II state. Preliminary histological analysis of lar- vae injected with recombinant protein suggested that the toxin primarily acted on the midgut of the insect. Finally, some of the common strategies used by the bac- terial pathogens of insects, animals, and plants are discussed. The availability of more than 100 microbial genomes, includ- ing the benign and pathogenic forms of closely related bacteria, is leading to an improved understanding of the common mech- anisms used by pathogenic bacteria to invade and survive in hosts. The current evidence suggests that bacterial pathogens of various hosts share many genes that aid virulence and survival (1, 2). The products of such genes are often called virulence factors and include factor