title: A Co-infection Model System and the Use of Chimeric Proteins to StudyChlamydiaInclusion Proteins Interaction.
abstract: Chlamydia trachomatisis an obligate intracellular bacterium associated with trachoma and sexually transmitted diseases. During its intracellular developmental cycle,Chlamydiaresides in a membrane bound compartment called the inclusion. A subset of Type III secreted effectors, the inclusion membrane proteins (Inc), are inserted into the inclusion membrane. Inc proteins are strategically positioned to promote inclusion interaction with host factors and organelles, a process required for bacterial replication, but little is known about Inc proteins function or host interacting partners. Moreover, it is unclear whether each Inc protein has a distinct function or if a subset of Inc proteins interacts with one another to perform their function. Here, we used IncD as a model to investigate Inc/Inc interaction in the context of Inc protein expression inC. trachomatis. We developed a co-infection model system to display different tagged Inc proteins on the surface of the same inclusion. We also designed chimeric Inc proteins to delineate domains important for interaction. We showed that IncD can self-interact and that the full-length protein is required for dimerization and/or oligomerization. Altogether our approach can be generalized to any Inc protein and will help to characterize the molecular mechanisms by whichChlamydiaInc proteins interact with themselves and/or host factors, eventually leading to a better understanding ofC. trachomatisinteraction with the mammalian host.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University New Haven, CT, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Han
authors.affiliation: Department of Microbiology, Immunology and Cancer Biology, University of Virginia Charlottesville, VA, USA.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Derré
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D001431:Bacteriological Techniques / Q000379:methods; D002692:Chlamydia trachomatis / Q000502:physiology*; D002479:Inclusion Bodies / Q000737:chemistry* / Q000382:microbiology*; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics; D025941:Protein Interaction Mapping; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins
keywords: Chlamydia; IncD; IncE; chimeric Inc protein; homo- and heterotypic interaction; inclusion membrane protein