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Journal of Bacteriology, March 2006, p. 2254-2261, Vol. 188, No. 6
0021-9193/06/$08.00+0     doi:10.1128/JB.188.6.2254-2261.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Photorhabdus Virulence Cassettes Confer Injectable Insecticidal Activity against the Wax Moth

G. Yang, A. J. Dowling, U. Gerike, R. H. ffrench-Constant, and N. R. Waterfield*

Center for Molecular Microbiology and Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, United Kingdom

Received 25 October 2005/ Accepted 19 December 2005

Two recently sequenced genomes of the insect-pathogenic bacterium Photorhabdus and a large Serratia entomophila plasmid, pADAP, have phage-related loci containing putative toxin effector genes, designated the "Photorhabdus virulence cassettes" (PVCs). In S. entomophila, the single plasmid PVC confers antifeeding activity on larvae of a beetle. Here, we show that recombinant Escherichia coli expressing PVC-containing cosmids from Photorhabdus has injectable insecticidal activity against larvae of the wax moth. Electron microscopy showed that the structure of the PVC products is similar to the structure of the antibacterial R-type pyocins. However, unlike these bacteriocins, the PVC products of Photorhabdus have no demonstrable antibacterial activity. Instead, injection of Photorhabdus PVC products destroys insect hemocytes, which undergo dramatic actin cytoskeleton condensation. Comparison of the genomic organizations of several PVCs showed that they have a conserved phage-like structure with a variable number of putative anti-insect effectors encoded at one end. Expression of these putative effectors directly inside cultured cells showed that they are capable of rearranging the actin cytoskeleton. Together, these data show that the PVCs are functional homologs of the S. entomophila antifeeding genes and encode physical structures that resemble bacteriocins. This raises the interesting hypothesis that the PVC products are bacteriocin-like but that they have been modified to attack eukaryotic host cells.

* Corresponding author. Mailing address: Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, United Kingdom. Phone: 44 1225 384292. Fax: 44 1225 386779. E-mail: bssnw{at}bath.ac.uk.

Journal of Bacteriology, March 2006, p. 2254-2261, Vol. 188, No. 6
0021-9193/06/$08.00+0     doi:10.1128/JB.188.6.2254-2261.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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