Chlamydia trachomatis Slc1 is a type III secretion chaperone that enhances the translocation of its invasion effector substrate TARP

Amanda J Brinkworth, Denise S Malcolm, António T Pedrosa, Katarzyna Roguska, Sevanna Shahbazian, James E Graham, Richard D Hayward, Rey A Carabeo

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Bacterial type III secretion system (T3SS) chaperones pilot substrates to the export apparatus in a secretion-competent state, and are consequently central to the translocation of effectors into target cells. Chlamydia trachomatis is a genetically intractable obligate intracellular pathogen that utilizes T3SS effectors to trigger its entry into mammalian cells. The only well-characterized T3SS effector is TARP (translocated actin recruitment protein), but its chaperone is unknown. Here we exploited a known structural signature to screen for putative type III secretion chaperones encoded within the C. trachomatis genome. Using bacterial two-hybrid, co-precipitation, cross-linking and size exclusion chromatography we show that Slc1 (SycE-like chaperone 1; CT043) specifically interacts with a 200-amino-acid residue N-terminal region of TARP (TARP¹¿²°°). Slc1 formed homodimers in vitro, as shown in cross-linking and gel filtration experiments. Biochemical analysis of an isolated Slc1-TARP¹¿²°° complex was consistent with a characteristic 2:1 chaperone-effector stoichiometry. Furthermore, Slc1 was co-immunoprecipitated with TARP from C. trachomatis elementary bodies. Also, coexpression of Slc1 specifically enhanced host cell translocation of TARP by a heterologous Yersinia enterocolitica T3SS. Taken together, we propose Slc1 as a chaperone of the C. trachomatis T3SS effector TARP.
Original languageEnglish
Pages (from-to)131-144
Number of pages14
JournalMolecular Microbiology
Volume82
Issue number1
Early online date1 Sep 2011
DOIs
Publication statusPublished - Oct 2011

Fingerprint

Chlamydia trachomatis
Actins
Proteins
Gel Chromatography
Yersinia enterocolitica
Protein C
Genome
Amino Acids

Keywords

  • bacterial proteins
  • sequence alignment
  • amino acid motifs
  • HeLa cells
  • humans
  • chlamydia infections
  • molecular sequence data
  • chlamydia trachomatis
  • molecular chaperones
  • amino acid sequence
  • protein binding
  • protein transport

Cite this

Brinkworth, A. J., Malcolm, D. S., Pedrosa, A. T., Roguska, K., Shahbazian, S., Graham, J. E., ... Carabeo, R. A. (2011). Chlamydia trachomatis Slc1 is a type III secretion chaperone that enhances the translocation of its invasion effector substrate TARP. Molecular Microbiology, 82(1), 131-144. https://doi.org/10.1111/j.1365-2958.2011.07802.x

Chlamydia trachomatis Slc1 is a type III secretion chaperone that enhances the translocation of its invasion effector substrate TARP. / Brinkworth, Amanda J; Malcolm, Denise S; Pedrosa, António T; Roguska, Katarzyna; Shahbazian, Sevanna; Graham, James E; Hayward, Richard D; Carabeo, Rey A.

In: Molecular Microbiology, Vol. 82, No. 1, 10.2011, p. 131-144.

Research output: Contribution to journalArticle

Brinkworth, AJ, Malcolm, DS, Pedrosa, AT, Roguska, K, Shahbazian, S, Graham, JE, Hayward, RD & Carabeo, RA 2011, 'Chlamydia trachomatis Slc1 is a type III secretion chaperone that enhances the translocation of its invasion effector substrate TARP', Molecular Microbiology, vol. 82, no. 1, pp. 131-144. https://doi.org/10.1111/j.1365-2958.2011.07802.x
Brinkworth, Amanda J ; Malcolm, Denise S ; Pedrosa, António T ; Roguska, Katarzyna ; Shahbazian, Sevanna ; Graham, James E ; Hayward, Richard D ; Carabeo, Rey A. / Chlamydia trachomatis Slc1 is a type III secretion chaperone that enhances the translocation of its invasion effector substrate TARP. In: Molecular Microbiology. 2011 ; Vol. 82, No. 1. pp. 131-144.
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abstract = "Bacterial type III secretion system (T3SS) chaperones pilot substrates to the export apparatus in a secretion-competent state, and are consequently central to the translocation of effectors into target cells. Chlamydia trachomatis is a genetically intractable obligate intracellular pathogen that utilizes T3SS effectors to trigger its entry into mammalian cells. The only well-characterized T3SS effector is TARP (translocated actin recruitment protein), but its chaperone is unknown. Here we exploited a known structural signature to screen for putative type III secretion chaperones encoded within the C. trachomatis genome. Using bacterial two-hybrid, co-precipitation, cross-linking and size exclusion chromatography we show that Slc1 (SycE-like chaperone 1; CT043) specifically interacts with a 200-amino-acid residue N-terminal region of TARP (TARP¹¿²°°). Slc1 formed homodimers in vitro, as shown in cross-linking and gel filtration experiments. Biochemical analysis of an isolated Slc1-TARP¹¿²°° complex was consistent with a characteristic 2:1 chaperone-effector stoichiometry. Furthermore, Slc1 was co-immunoprecipitated with TARP from C. trachomatis elementary bodies. Also, coexpression of Slc1 specifically enhanced host cell translocation of TARP by a heterologous Yersinia enterocolitica T3SS. Taken together, we propose Slc1 as a chaperone of the C. trachomatis T3SS effector TARP.",
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AU - Roguska, Katarzyna

AU - Shahbazian, Sevanna

AU - Graham, James E

AU - Hayward, Richard D

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