Chlamydia trachomatis tmea directly activates N-WASP to promote actin polymerization and functions synergistically with TarP during invasion

Gabrielle Keb, Joshua Ferrell, Kaylyn R. Scanlon, Travis J. Jewett, Kenneth A. Fields

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Chlamydia trachomatis is a medically significant human pathogen and is an epithelial-tropic obligate intracellular parasite. Invasion of nonprofessional phago-cytes represents a crucial step in the infection process and has likely promoted the evolution of a redundant mechanism and routes of entry. Like many other viral and invasive bacterial pathogens, manipulation of the host cell cytoskeleton represents a focal point in Chlamydia entry. The advent of genetic techniques in C. trachomatis, such as creation of complete gene deletions via fluorescence-reported allelic exchange mutagenesis (FRAEM), is providing important tools to unravel the contributions of bacterial factors in these complex pathways. The type III secretion chaperone Slc1 directs delivery of at least four effectors during the invasion process. Two of these, TarP and TmeA, have been associated with manipulation of actin networks and are essential for normal levels of invasion. The functions of TarP are well established, whereas TmeA is less well characterized. We leverage chlamydial genetics and proximity labeling here to provide evidence that TmeA directly targets host N-WASP to promote Arp2/3-depend-ent actin polymerization. Our work also shows that TmeA and TarP influence separate, yet synergistic pathways to accomplish chlamydial entry. These data further support an appreciation that a pathogen, confined by a reductionist genome, retains the ability to commit considerable resources to accomplish bottle-neck steps during the infection process. IMPORTANCE The increasing genetic tractability of Chlamydia trachomatis is accelerat-ing the ability to characterize the unique infection biology of this obligate intracellular parasite. These efforts are leading to a greater understanding of the molecular events associated with key virulence requirements. Manipulation of the host actin cy-toskeleton plays a pivotal role throughout Chlamydia infection, yet a thorough understanding of the molecular mechanisms initiating and orchestrating actin rear-rangements has lagged. Our work highlights the application of genetic manipulation to address open questions regarding chlamydial invasion, a process essential to sur-vival. We provide definitive insight regarding the role of the type III secreted effector TmeA and how that activity relates to another prominent effector, TarP. In addition, our data implicate at least one source that contributes to the functional divergence of entry mechanisms among chlamydial species.

Original languageEnglish
Article numbere02861-20
Pages (from-to)1-18
Number of pages18
JournalmBio
Volume12
Issue number1
DOIs
StatePublished - 2021

Bibliographical note

Funding Information:
We thank Mollie Jewett and members of the Fields and Travis Jewett lab for critical reading of the manuscript. We also thank Raphael Valdivia for the kind gift of TepP-specific antibodies, and Maria Clouse and Robert Hayman for excellent technical assistance. Mass spectroscopy analyses were performed by the University of Kentucky Proteomics Core Facility, and we acknowledge the helpful expertise of Jing Chen and Haining Zhu. This work was supported by Public Health Service grants from the National Institutes of Health, NIAID, to K.A.F. (AI065530), T.J.J. (AI139242), and G.K. (AI147417).

Funding Information:
This work was supported by Public Health Service grants from the National Institutes of Health, NIAID, to K.A.F. (AI065530), T.J.J. (AI139242), and G.K. (AI147417).

Publisher Copyright:
© 2021 Keb et al.

Keywords

  • FLAEM
  • FRAEM
  • N-WASP
  • Type III secretion

ASJC Scopus subject areas

  • Microbiology
  • Virology

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