CteG is a Chlamydia trachomatis effector protein that associates with the Golgi complex of infected host cells

Sara V. Pais, Charlotte E. Key, Vítor Borges, Inês S. Pereira, João Paulo Gomes, Derek J. Fisher, Luís Jaime Mota

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Abstract

Chlamydia trachomatis is a bacterial pathogen causing ocular and genital infections in humans. C. trachomatis multiplies exclusively inside host cells within a characteristic vacuole, from where it manipulates host cells by injecting them with type III secretion effector proteins. Here, we identified CteG as the first C. trachomatiseffector associated with the Golgi. For this, C. trachomatis strains expressing candidate effectors fused to a double hemagglutinin (2HA) tag were constructed. Then, among these strains, immunofluorescence microscopy revealed that CteG-2HA was delivered into the cytoplasm of infected cells. Between 16–20 h post-infection, CteG-2HA mostly associated with the Golgi; however, CteG-2HA also appeared at the host cell plasma membrane, and at 30 or 40 h post-infection this was its predominant localization. This change in the main localization of CteG-2HA was independent of intact microfilaments or microtubules. Ectopic expression of different regions of CteG (656 amino acid residues) in uninfected cells revealed that its first 100 residues contain a Golgi targeting region. Although a C. trachomatis cteG mutant did not display a defect in intracellular multiplication, CteG induced a vacuolar protein sorting defect when expressed in Saccharomyces cerevisiae. This suggested that CteG might function by subverting host cell vesicular transport.

Original languageEnglish
Article number6133
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 1 Dec 2019

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Chlamydia trachomatis
Golgi Apparatus
Proteins
Cell Membrane
Eye Infections
Hemagglutinins
Protein Transport
Vacuoles
Infection
Actin Cytoskeleton
Fluorescence Microscopy
Microtubules
Saccharomyces cerevisiae
Cytoplasm
Amino Acids

Cite this

@article{60fbcabdee604843aea34133fe21be47,
title = "CteG is a Chlamydia trachomatis effector protein that associates with the Golgi complex of infected host cells",
abstract = "Chlamydia trachomatis is a bacterial pathogen causing ocular and genital infections in humans. C. trachomatis multiplies exclusively inside host cells within a characteristic vacuole, from where it manipulates host cells by injecting them with type III secretion effector proteins. Here, we identified CteG as the first C. trachomatiseffector associated with the Golgi. For this, C. trachomatis strains expressing candidate effectors fused to a double hemagglutinin (2HA) tag were constructed. Then, among these strains, immunofluorescence microscopy revealed that CteG-2HA was delivered into the cytoplasm of infected cells. Between 16–20 h post-infection, CteG-2HA mostly associated with the Golgi; however, CteG-2HA also appeared at the host cell plasma membrane, and at 30 or 40 h post-infection this was its predominant localization. This change in the main localization of CteG-2HA was independent of intact microfilaments or microtubules. Ectopic expression of different regions of CteG (656 amino acid residues) in uninfected cells revealed that its first 100 residues contain a Golgi targeting region. Although a C. trachomatis cteG mutant did not display a defect in intracellular multiplication, CteG induced a vacuolar protein sorting defect when expressed in Saccharomyces cerevisiae. This suggested that CteG might function by subverting host cell vesicular transport.",
author = "Pais, {Sara V.} and Key, {Charlotte E.} and V{\'i}tor Borges and Pereira, {In{\^e}s S.} and Gomes, {Jo{\~a}o Paulo} and Fisher, {Derek J.} and Mota, {Lu{\'i}s Jaime}",
note = "This work was supported by Fundacao para a Ciencia e a Tecnologia (FCT/MCTES) through grants PTDC/IMI-MIC/1300/2014 and PTDC/BIA-MIC/28503/2017, and by the Applied Molecular Biosciences Unit (UCIBIO), which is financed by national funds from FCT/MCTES (UID/Multi/04378/2019) and co-financed by the European Regional Development Fund (ERDF) under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728). SVP was supported by PhD fellowship PD/BD/52210/2013 within the scope of the PhD program Molecular Biosciences (PD/00133/2012) funded by FCT/MCTES. ISP was supported by PhD fellowship SFRH/BD/129756/2017. Work by DJF was funded by grant R21AI115238 from the National Institutes of Health.",
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CteG is a Chlamydia trachomatis effector protein that associates with the Golgi complex of infected host cells. / Pais, Sara V.; Key, Charlotte E.; Borges, Vítor; Pereira, Inês S.; Gomes, João Paulo; Fisher, Derek J.; Mota, Luís Jaime.

In: Scientific Reports, Vol. 9, No. 1, 6133, 01.12.2019.

Research output: Contribution to journalArticle

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T1 - CteG is a Chlamydia trachomatis effector protein that associates with the Golgi complex of infected host cells

AU - Pais, Sara V.

AU - Key, Charlotte E.

AU - Borges, Vítor

AU - Pereira, Inês S.

AU - Gomes, João Paulo

AU - Fisher, Derek J.

AU - Mota, Luís Jaime

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PY - 2019/12/1

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N2 - Chlamydia trachomatis is a bacterial pathogen causing ocular and genital infections in humans. C. trachomatis multiplies exclusively inside host cells within a characteristic vacuole, from where it manipulates host cells by injecting them with type III secretion effector proteins. Here, we identified CteG as the first C. trachomatiseffector associated with the Golgi. For this, C. trachomatis strains expressing candidate effectors fused to a double hemagglutinin (2HA) tag were constructed. Then, among these strains, immunofluorescence microscopy revealed that CteG-2HA was delivered into the cytoplasm of infected cells. Between 16–20 h post-infection, CteG-2HA mostly associated with the Golgi; however, CteG-2HA also appeared at the host cell plasma membrane, and at 30 or 40 h post-infection this was its predominant localization. This change in the main localization of CteG-2HA was independent of intact microfilaments or microtubules. Ectopic expression of different regions of CteG (656 amino acid residues) in uninfected cells revealed that its first 100 residues contain a Golgi targeting region. Although a C. trachomatis cteG mutant did not display a defect in intracellular multiplication, CteG induced a vacuolar protein sorting defect when expressed in Saccharomyces cerevisiae. This suggested that CteG might function by subverting host cell vesicular transport.

AB - Chlamydia trachomatis is a bacterial pathogen causing ocular and genital infections in humans. C. trachomatis multiplies exclusively inside host cells within a characteristic vacuole, from where it manipulates host cells by injecting them with type III secretion effector proteins. Here, we identified CteG as the first C. trachomatiseffector associated with the Golgi. For this, C. trachomatis strains expressing candidate effectors fused to a double hemagglutinin (2HA) tag were constructed. Then, among these strains, immunofluorescence microscopy revealed that CteG-2HA was delivered into the cytoplasm of infected cells. Between 16–20 h post-infection, CteG-2HA mostly associated with the Golgi; however, CteG-2HA also appeared at the host cell plasma membrane, and at 30 or 40 h post-infection this was its predominant localization. This change in the main localization of CteG-2HA was independent of intact microfilaments or microtubules. Ectopic expression of different regions of CteG (656 amino acid residues) in uninfected cells revealed that its first 100 residues contain a Golgi targeting region. Although a C. trachomatis cteG mutant did not display a defect in intracellular multiplication, CteG induced a vacuolar protein sorting defect when expressed in Saccharomyces cerevisiae. This suggested that CteG might function by subverting host cell vesicular transport.

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