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
The translocated actin recruiting phosphoprotein (Tarp) is a multidomain type III secreted effector used by 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 , we speculated as to what degree Tarp...
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Published in | Infection and immunity Vol. 88; no. 5 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
American Society for Microbiology
20.04.2020
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Subjects | |
Online Access | Get full text |
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Summary: | The translocated actin recruiting phosphoprotein (Tarp) is a multidomain type III secreted effector used by
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
, we speculated as to what degree Tarp function contributes to
's ability to parasitize mammalian host cells. To address this question, we generated a complete
deletion mutant using the fluorescence-reported allelic exchange mutagenesis (FRAEM) technique and complemented the mutant in
with wild-type
or mutant
alleles engineered to harbor in-frame domain deletions. We provide evidence for the significant role of Tarp in
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
entry into HeLa cells resulted in host cell shape changes, whereas the
mutant did not. Finally, using a novel
complementation approach,
lacking
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
virulence factor. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Citation Ghosh S, Ruelke EA, Ferrell JC, Bodero MD, Fields KA, Jewett TJ. 2020. 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. Infect Immun 88:e00841-19. https://doi.org/10.1128/IAI.00841-19. |
ISSN: | 0019-9567 1098-5522 |
DOI: | 10.1128/IAI.00841-19 |