Abstract
Development of approaches to genetically manipulate Chlamydia is fostering important advances in understanding pathogenesis. Fluorescence-reported allelic exchange mutagenesis (FRAEM) now enables the complete deletion of specific genes in C. trachomatis L2. We have leveraged this technology to delete the coding sequences for a known type III effector. The evidence provided here indicates that CT694/CTL0063 is a virulence protein involved in chlamydial invasion. Based on our findings, we designate the gene product corresponding to ct694-ctl0063 translocated membrane-associated effector A (TmeA). Deletion of tmeA did not impact development of intracellular chlamydiae. However, the absence of TmeA manifested as a decrease in infectivity in both tissue culture and murine infection models. The in vitro defect was reflected by impaired invasion of host cells. TmeA binds human AHNAK, and we demonstrate here that AHNAK is transiently recruited by invading chlamydiae. TmeA, however, is not required for endogenous AHNAK recruitment. TmeA also impairs AHNAK-dependent actin bundling activity. This TmeA-mediated effect likely does not explain impaired invasion displayed by the tmeA strain of Chlamydia, since AHNAK-deficient cells revealed no invasion phenotype. Overall, our data indicate the efficacy of FRAEM and reveal a role of TmeA during chlamydial invasion that manifests independently of effects on AHNAK.
Document Type
Article
Publication Date
12-2017
Digital Object Identifier (DOI)
https://doi.org/10.1128/IAI.00640-17
Funding Information
This work was supported by Public Health Service grants from the National Institutes of Health, NIAID (AI065530 and AI124649), to K.A.F.
Related Content
Supplemental material for this article may be found at https://doi.org/10.1128/IAI.00640-17.
Repository Citation
McKuen, M. J.; Mueller, Konrad E.; Bae, Y. S.; and Fields, Kenneth A., "Fluorescence-Reported Allelic Exchange Mutagenesis Reveals a Role for Chlamydia trachomatis TmeA in Invasion That Is Independent of Host AHNAK" (2017). Microbiology, Immunology, and Molecular Genetics Faculty Publications. 101.
https://uknowledge.uky.edu/microbio_facpub/101
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Notes/Citation Information
Published in Infection and Immunity, v. 85, issue 12, e00640-17, p. 1-18.
Copyright © 2017 American Society for Microbiology. All Rights Reserved.
The copyright holder has granted the permission for posting the article here.