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Published on: October 9, 2010
Mimicry of a G protein mutation by pertussis toxin expression in transgenic Caenorhabditis elegans
1Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California 94305-5124, USA. cdarby@stanford.edu
Abstract:
Pathogens produce virulence factors that interact directly with host molecules, but in many cases the host targets are unknown. The genetic and molecular identification of these orphan targets is often not feasible with mammalian experimental models. However, a substantial number of known targets are molecules and pathways that are conserved among eukaryotes, and therefore the use of nonmammalian model hosts to identify orphan targets may prove useful. To demonstrate the feasibility of this approach, we transformed the nematode Caenorhabditis elegans with a gene encoding the catalytic subunit of pertussis toxin (PTX), which in mammals inactivates G(o/i)alpha proteins. Expression of PTX in C. elegans produced phenotypes almost identical to those of a null mutation in the nematode gene encoding G(o/i)alpha. Furthermore, PTX suppressed the phenotype of a constitutively active form of nematode G(o/i)alpha protein. These results indicate that PTX is functional in nematodes and acts specifically on the C. elegans homologue of the mammalian target.
Insights
This study shows that pertussis toxin (PTX) can be used in the nematode Caenorhabditis elegans to identify unknown pathogen targets. This nonmammalian model approach successfully identified a conserved G(o/i)alpha protein as a PTX target.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Pathogen virulence factors target host molecules, but these targets are often unknown.
- Identifying these "orphan targets" is challenging in mammalian models.
- Eukaryotic-conserved molecules and pathways offer potential targets for nonmammalian models.
Purpose of the Study:
- To demonstrate the feasibility of using the nematode Caenorhabditis elegans to identify unknown pathogen targets.
- To validate the use of pertussis toxin (PTX) as a tool in a nonmammalian host.
Main Methods:
- Transformed Caenorhabditis elegans with the gene for the catalytic subunit of pertussis toxin (PTX).
- Assessed PTX function by observing phenotypes related to G(o/i)alpha protein activity.
- Compared PTX-induced phenotypes with those of a null mutation in the nematode G(o/i)alpha gene.
Main Results:
- Expression of PTX in C. elegans resulted in phenotypes nearly identical to a null mutation in the nematode G(o/i)alpha gene.
- PTX effectively suppressed the phenotype of a constitutively active form of nematode G(o/i)alpha protein.
- Demonstrated PTX functionality and specificity towards the C. elegans homologue of mammalian G(o/i)alpha.
Conclusions:
- Pertussis toxin is functional in the nematode C. elegans.
- C. elegans can serve as a model host for identifying pathogen targets conserved across eukaryotes.
- This approach facilitates the study of pathogen-host interactions and the identification of novel therapeutic targets.
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