Photorhabdus luminescens W-14 Insecticidal Activity Consists of at Least Two Similar but Distinct Proteins

Guo, Lining; Fatig, Raymond; Orr, Gregory L.; Schafer, Barry W.; Strickland, James A.; Sukhapinda, Kitisri; Woodsworth, Aaron T.; Petell, James K.

doi:10.1074/jbc.274.14.9836

Cited by 63 publications

(44 citation statements)

References 37 publications

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“…From earlier studies, it has been suggested that class A proteins harbor the cytotoxic effects of the Tc toxins, whereas class B and C proteins modulate and enhance the toxicity of class A proteins (16). However, recently, we elucidated the molecular mechanism of the Photorhabdus luminescens Tc complex, which consists of the class A protein TcdA1, the class B protein TcdB2, and the class C protein TccC3 or TccC5 (17).…”

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confidence: 99%

Insecticidal Toxin Complex Proteins from Xenorhabdus nematophilus

Sheets¹,

Hey²,

Fencil³

et al. 2011

Journal of Biological Chemistry

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Toxin complexes from Xenorhabdus and Photorhabdus spp. bacteria represent novel insecticidal proteins. We purified a native toxin complex (toxin complex 1) from Xenorhabdus nematophilus. The toxin complex is composed of three different proteins, XptA2, XptB1, and XptC1, representing products from class A, B, and C toxin complex genes, respectively. We showed that recombinant XptA2 and co-produced recombinant XptB1 and XptC1 bind together with a 4:1:1 stoichiometry. XptA2 forms a tetramer of ϳ1,120 kDa that bound to solubilized insect brush border membranes and induced pore formation in black lipid membranes. Co-expressed XptB1 and XptC1 form a tight 1:1 binary complex where XptC1 is C-terminally truncated, resulting in a 77-kDa protein. The ϳ30-kDa C-terminally cleaved portion of XptC1 apparently only loosely associates with this binary complex. XptA2 had only modest oral toxicity against lepidopteran insects but as a complex with co-produced XptB1 and XptC1 had high levels of insecticidal activity. Addition of co-expressed class B (TcdB2) and class C (TccC3) proteins from Photorhabdus luminescens to the Xenorhabdus XptA2 protein resulted in formation of a hybrid toxin complex protein with the same 4:1:1 stoichiometry as the native Xenorhabdus toxin complex 1. This hybrid toxin complex, like the native toxin complex, was highly active against insects.Xenorhabdus and Photorhabdus spp. are two bacterial genera belonging to the family Enterobacteriaceae, known to be associated with entomopathogenic nematodes (1-4) These bacteria represent potential sources for new genes encoding potent insecticidal toxins that could be put into plants as alternatives to Bacillus thuringiensis genes (5). Gene sequence analysis of Xenorhabdus and Photorhabdus bacteria show that these organisms contain a family of related toxin complex (tc) 2 genes located at different loci (6 -9). The toxin complexes are composed of three different classes of protein components, which, according to ffrench-Constant et al. (10,11), can be categorized as class A, B, and C proteins based upon sequence similarity and size. Class A proteins are very large, having a molecular mass of ϳ280 kDa. Class B proteins are ϳ170 kDa, and class C proteins are ϳ110 kDa. There are many different varieties of class A, B, and C proteins in both Gram-negative and Gram-positive bacteria (12-15).From earlier studies, it has been suggested that class A proteins harbor the cytotoxic effects of the Tc toxins, whereas class B and C proteins modulate and enhance the toxicity of class A proteins (16). However, recently, we elucidated the molecular mechanism of the Photorhabdus luminescens Tc complex, which consists of the class A protein TcdA1, the class B protein TcdB2, and the class C protein TccC3 or TccC5 (17). These studies revealed that the class C proteins harbor the biological activity. It was shown that TccC3 and TccC5 are ADP-ribosyltransferases, which target the actin cytoskeleton by modification of actin and Rho GTPases, respectively (17). Moreover, these studies su...

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confidence: 99%

Insecticidal Toxin Complex Proteins from Xenorhabdus nematophilus

Sheets¹,

Hey²,

Fencil³

et al. 2011

Journal of Biological Chemistry

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“…Other toxin proteins isolated and characterized from bacteria related to Xenorhabdus are the Tc toxins from P. luminescens strain W14 (17)(18)(19), a 39-kDa toxin from X. nematophila (20), the large Xin toxin from X. nematophila strain BJ (21), and the PhlA hemolysin from P. luminescens strain TT01 (22). Of these, only the Tc toxins have been studied in any detail.…”

Section: Discussionmentioning

confidence: 99%

“…The PhlA hemolysin was shown not to be a major virulence determinant (22). The Tc toxins from P. luminescens W14 form a large protein complex that is toxic to insects by either ingestion or injection (17)(18)(19). The high molecular mass complex consists of about 10 polypeptides, ranging in size from 30 to 200 kDa, which is secreted from the bacteria and requires proteolytic processing for activity.…”

Section: Discussionmentioning

confidence: 99%

“…Searches of the non-redundant nucleotide and protein data bases (released Oct. 17,2003) were performed using the various BLAST programs (version 2.2.6) available via NCBI (www.ncbi.nlm.nih.gov/BLAST/). Motif and fold recognition analysis of the protein sequence was performed using programs such as InterProScan (www.ebi.ac.uk/interpro/, July 22, 2003 (28)), PANAL (mgd.ahc.umn.edu/panal/ (29)), Genequiz (jura.ebi.ac.uk:8765/extgenequiz/ (30)), and the Structure Prediction Metaserver (bioinfo.pl/ Meta/pdb-test.pl/ (31)).…”

Section: Methodsmentioning

confidence: 99%

“…Analysis of the genome of P. luminescens identified more predicted toxin genes than any other bacteria sequenced to date, including potential gene products with homology to hemolysin A, chitinase, Rtx (repeats-in-toxin)-like toxin, and ␦-endotoxin (5). The only toxins studied in detail are the Tc toxins from P. luminescens strain W14 (17)(18)(19), although a small amount of work has also been done on a 39-kDa toxin from X. nematophila (20), the large Xin toxin from X. nematophila strain BJ (21), and the PhlA hemolysin from P. luminescens strain TT01 (22). This paper describes the identification, purification, and characterization of a novel insect toxin from X. nematophila strain A24.…”

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

Brown

Cao

Hines

et al. 2004

Journal of Biological Chemistry

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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 bioinformatic analysis suggested that the toxin was a novel protein, 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 larvae 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 bacterial pathogens of insects, animals, and plants are discussed.

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The Genera Photorhabdus and Xenorhabdus

Boemare¹,

Akhurst²

2006

The Prokaryotes

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Photorhabdus luminescens W-14 Insecticidal Activity Consists of at Least Two Similar but Distinct Proteins

Cited by 63 publications

References 37 publications

Insecticidal Toxin Complex Proteins from Xenorhabdus nematophilus

Insecticidal Toxin Complex Proteins from Xenorhabdus nematophilus

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

The Genera Photorhabdus and Xenorhabdus

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