Chlamydia Trachomatis Inclusion Membrane Protein CTL0478 Interaction with Flightless I Homolog

Abstract

Chlamydia trachomatis causes a range of infections such as blinding trachoma and urogenital infections leading to serious complications. Inside the host cells, it lives in a parasitophorous vacuole called inclusion and secretes various effectors to manipulate host’s cellular processes. CTL0478 is an inclusion membrane protein (Inc) shown to co-purify with flightless I homolog (FLII) in a global Inc-human interactome study. FLII is known to regulate inflammation pathways. This study aims to characterize CTL0478-FLII interaction. In silico analysis of CTL0478 revealed a leucine zipper (LZ) within coiled coil in the C-terminal. Coiled coil is also found in host proteins interacting with FLII. C. muridarum contains a CTL0478 homolog with conserved leucine residues in LZ. FLII recruitment to C. trachomatis L2 inclusion was confirmed by Immunofluorescence. FLII recruitment was also seen in C. trachomatis serovar B and C. muridarum suggesting an overall conserved mechanism. HeLa cell line expressing CTL0478-GFP was generated for use in directed pull-down assays. Further, CTL0478 was cloned into chlamydial shuttle vector pBOMB-tet-mCherry which will be used to generate C. trachomatis L2 expressing flag tagged CTL0478 for co-localization and pull down assays.