A Coinfection Model to Evaluate Chlamydia Inc Protein Interactions

Abstract

Chlamydia trachomatis resides and replicates within a membranous vacuole, termed the inclusion. A group of Type III secreted effector proteins, the inclusion membrane proteins (Inc), are embedded within the inclusion membrane and facilitate the interaction of the inclusion with host cell organelles. These interactions are vital for bacterial replication and allow for the acquisition of essential nutrients from the host cell. However, it is not known if Inc proteins function independently or require interactions with other Inc proteins to function. This chapter describes a system to test the homotypic/heterotypic interactions of Inc proteins through the coinfection of Chlamydia strains expressing differently tagged inclusion membrane proteins. Our approach takes advantage of the natural homotypic fusion of inclusions and allows for the study of Inc protein interactions when they are embedded within the inclusion membrane.