Fluorescence-reported allelic exchange mutagenesis-mediated gene deletion indicates a requirement for chlamydia trachomatis tarp during in vivo infectivity and reveals a specific role for the C terminus during cellular invasion

Susmita Ghosh, Elizabeth A. Ruelke, Joshua C. Ferrell, Maria D. Bodero, Kenneth A. Fields, Travis J. Jewett

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

The translocated actin recruiting phosphoprotein (Tarp) is a multidomain type III secreted effector used by Chlamydia trachomatis. In aggregate, existing data suggest a role of this effector in initiating new infections. As new genetic tools began to emerge to study chlamydial genes in vivo, we speculated as to what degree Tarp function contributes to Chlamydia's ability to parasitize mammalian host cells. To address this question, we generated a complete tarP deletion mutant using the fluorescence-reported allelic exchange mutagenesis (FRAEM) technique and complemented the mutant in trans with wild-type tarP or mutant tarP alleles engineered to harbor in-frame domain deletions. We provide evidence for the significant role of Tarp in C. trachomatis invasion of host cells. Complementation studies indicate that the C-terminal filamentous actin (F-actin)-binding domains are responsible for Tarp-mediated invasion efficiency. Wild-type C. trachomatis entry into HeLa cells resulted in host cell shape changes, whereas the tarP mutant did not. Finally, using a novel cis complementation approach, C. trachomatis lacking tarP demonstrated significant attenuation in a murine genital tract infection model. Together, these data provide definitive genetic evidence for the critical role of the Tarp F-actin-binding domains in host cell invasion and for the Tarp effector as a bona fide C. trachomatis virulence factor.

Original languageEnglish
Article numbere00841-19
JournalInfection and Immunity
Volume88
Issue number5
DOIs
StatePublished - May 1 2020

Bibliographical note

Publisher Copyright:
© 2020 American Society for Microbiology. All Rights Reserved.

Keywords

  • Actin
  • Actin cytoskeleton
  • Chlamydia trachomatis
  • Cytoskeleton
  • Effector
  • Effector functions
  • Tarp

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

Fingerprint

Dive into the research topics of 'Fluorescence-reported allelic exchange mutagenesis-mediated gene deletion indicates a requirement for chlamydia trachomatis tarp during in vivo infectivity and reveals a specific role for the C terminus during cellular invasion'. Together they form a unique fingerprint.

Cite this