A chlamydial type III translocated protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin

D R Clifton, K A Fields, S S Grieshaber, C A Dooley, E R Fischer, D J Mead, R A Carabeo, T Hackstadt

Research output: Contribution to journalArticle

276 Citations (Scopus)

Abstract

The obligate intracellular bacterium Chlamydia trachomatis rapidly induces its own entry into host cells. Initial attachment is mediated by electrostatic interactions to heparan sulfate moieties on the host cell, followed by irreversible binding to an unknown secondary receptor. This secondary binding leads to the recruitment of actin to the site of attachment, formation of an actin-rich, pedestal-like structure, and finally internalization of the bacteria. How chlamydiae induce this process is unknown. We have identified a high-molecular-mass tyrosine-phosphorylated protein that is rapidly phosphorylated on attachment to the host cell. Immunoelectron microscopy studies revealed that this tyrosine-phosphorylated protein is localized to the cytoplasmic face of the plasma membrane at the site of attachment of surface-associated chlamydiae. The phosphoprotein was isolated by immunoprecipitation with the antiphosphotyrosine antibody 4G10 and identified as the chlamydial protein CT456, a hypothetical protein with unknown function. The chlamydial protein (Tarp) appears to be translocated into the host cell by type III secretion because it is exported in a Yersinia heterologous expression assay. Phosphotyrosine signaling across the plasma membrane preceded the recruitment of actin to the site of chlamydial attachment and may represent the initial signal transduced from pathogen to the host cell. These results suggest that C. trachomatis internalization is mediated by a chlamydial type III-secreted effector protein.
Original languageEnglish
Pages (from-to)10166-71
Number of pages6
JournalPNAS
Volume101
Issue number27
DOIs
Publication statusPublished - 6 Jul 2004

Fingerprint

Tyrosine
Actins
Proteins
Chlamydia
Chlamydia trachomatis
Cell Membrane
Bacteria
Yersinia
Phosphotyrosine
Heparitin Sulfate
Immunoelectron Microscopy
Phosphoproteins
Static Electricity
Immunoprecipitation
Antibodies

Keywords

  • Animals
  • COS Cells
  • HeLa Cells
  • Humans
  • Tyrosine
  • Amino Acid Sequence
  • Bacterial Proteins
  • Phosphorylation
  • Transfection
  • Actins
  • Molecular Sequence Data
  • Chlamydia
  • Protein Transport

Cite this

Clifton, D. R., Fields, K. A., Grieshaber, S. S., Dooley, C. A., Fischer, E. R., Mead, D. J., ... Hackstadt, T. (2004). A chlamydial type III translocated protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin. PNAS, 101(27), 10166-71. https://doi.org/10.1073/pnas.0402829101

A chlamydial type III translocated protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin. / Clifton, D R; Fields, K A; Grieshaber, S S; Dooley, C A; Fischer, E R; Mead, D J; Carabeo, R A; Hackstadt, T.

In: PNAS, Vol. 101, No. 27, 06.07.2004, p. 10166-71.

Research output: Contribution to journalArticle

Clifton, DR, Fields, KA, Grieshaber, SS, Dooley, CA, Fischer, ER, Mead, DJ, Carabeo, RA & Hackstadt, T 2004, 'A chlamydial type III translocated protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin', PNAS, vol. 101, no. 27, pp. 10166-71. https://doi.org/10.1073/pnas.0402829101
Clifton, D R ; Fields, K A ; Grieshaber, S S ; Dooley, C A ; Fischer, E R ; Mead, D J ; Carabeo, R A ; Hackstadt, T. / A chlamydial type III translocated protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin. In: PNAS. 2004 ; Vol. 101, No. 27. pp. 10166-71.
@article{25d06f7474844dac8f6ee31504d2781a,
title = "A chlamydial type III translocated protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin",
abstract = "The obligate intracellular bacterium Chlamydia trachomatis rapidly induces its own entry into host cells. Initial attachment is mediated by electrostatic interactions to heparan sulfate moieties on the host cell, followed by irreversible binding to an unknown secondary receptor. This secondary binding leads to the recruitment of actin to the site of attachment, formation of an actin-rich, pedestal-like structure, and finally internalization of the bacteria. How chlamydiae induce this process is unknown. We have identified a high-molecular-mass tyrosine-phosphorylated protein that is rapidly phosphorylated on attachment to the host cell. Immunoelectron microscopy studies revealed that this tyrosine-phosphorylated protein is localized to the cytoplasmic face of the plasma membrane at the site of attachment of surface-associated chlamydiae. The phosphoprotein was isolated by immunoprecipitation with the antiphosphotyrosine antibody 4G10 and identified as the chlamydial protein CT456, a hypothetical protein with unknown function. The chlamydial protein (Tarp) appears to be translocated into the host cell by type III secretion because it is exported in a Yersinia heterologous expression assay. Phosphotyrosine signaling across the plasma membrane preceded the recruitment of actin to the site of chlamydial attachment and may represent the initial signal transduced from pathogen to the host cell. These results suggest that C. trachomatis internalization is mediated by a chlamydial type III-secreted effector protein.",
keywords = "Animals, COS Cells, HeLa Cells, Humans, Tyrosine, Amino Acid Sequence, Bacterial Proteins, Phosphorylation, Transfection, Actins, Molecular Sequence Data, Chlamydia, Protein Transport",
author = "Clifton, {D R} and Fields, {K A} and Grieshaber, {S S} and Dooley, {C A} and Fischer, {E R} and Mead, {D J} and Carabeo, {R A} and T Hackstadt",
year = "2004",
month = "7",
day = "6",
doi = "10.1073/pnas.0402829101",
language = "English",
volume = "101",
pages = "10166--71",
journal = "PNAS",
issn = "0027-8424",
publisher = "NATL ACAD SCIENCES",
number = "27",

}

TY - JOUR

T1 - A chlamydial type III translocated protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin

AU - Clifton, D R

AU - Fields, K A

AU - Grieshaber, S S

AU - Dooley, C A

AU - Fischer, E R

AU - Mead, D J

AU - Carabeo, R A

AU - Hackstadt, T

PY - 2004/7/6

Y1 - 2004/7/6

N2 - The obligate intracellular bacterium Chlamydia trachomatis rapidly induces its own entry into host cells. Initial attachment is mediated by electrostatic interactions to heparan sulfate moieties on the host cell, followed by irreversible binding to an unknown secondary receptor. This secondary binding leads to the recruitment of actin to the site of attachment, formation of an actin-rich, pedestal-like structure, and finally internalization of the bacteria. How chlamydiae induce this process is unknown. We have identified a high-molecular-mass tyrosine-phosphorylated protein that is rapidly phosphorylated on attachment to the host cell. Immunoelectron microscopy studies revealed that this tyrosine-phosphorylated protein is localized to the cytoplasmic face of the plasma membrane at the site of attachment of surface-associated chlamydiae. The phosphoprotein was isolated by immunoprecipitation with the antiphosphotyrosine antibody 4G10 and identified as the chlamydial protein CT456, a hypothetical protein with unknown function. The chlamydial protein (Tarp) appears to be translocated into the host cell by type III secretion because it is exported in a Yersinia heterologous expression assay. Phosphotyrosine signaling across the plasma membrane preceded the recruitment of actin to the site of chlamydial attachment and may represent the initial signal transduced from pathogen to the host cell. These results suggest that C. trachomatis internalization is mediated by a chlamydial type III-secreted effector protein.

AB - The obligate intracellular bacterium Chlamydia trachomatis rapidly induces its own entry into host cells. Initial attachment is mediated by electrostatic interactions to heparan sulfate moieties on the host cell, followed by irreversible binding to an unknown secondary receptor. This secondary binding leads to the recruitment of actin to the site of attachment, formation of an actin-rich, pedestal-like structure, and finally internalization of the bacteria. How chlamydiae induce this process is unknown. We have identified a high-molecular-mass tyrosine-phosphorylated protein that is rapidly phosphorylated on attachment to the host cell. Immunoelectron microscopy studies revealed that this tyrosine-phosphorylated protein is localized to the cytoplasmic face of the plasma membrane at the site of attachment of surface-associated chlamydiae. The phosphoprotein was isolated by immunoprecipitation with the antiphosphotyrosine antibody 4G10 and identified as the chlamydial protein CT456, a hypothetical protein with unknown function. The chlamydial protein (Tarp) appears to be translocated into the host cell by type III secretion because it is exported in a Yersinia heterologous expression assay. Phosphotyrosine signaling across the plasma membrane preceded the recruitment of actin to the site of chlamydial attachment and may represent the initial signal transduced from pathogen to the host cell. These results suggest that C. trachomatis internalization is mediated by a chlamydial type III-secreted effector protein.

KW - Animals

KW - COS Cells

KW - HeLa Cells

KW - Humans

KW - Tyrosine

KW - Amino Acid Sequence

KW - Bacterial Proteins

KW - Phosphorylation

KW - Transfection

KW - Actins

KW - Molecular Sequence Data

KW - Chlamydia

KW - Protein Transport

U2 - 10.1073/pnas.0402829101

DO - 10.1073/pnas.0402829101

M3 - Article

C2 - 15199184

VL - 101

SP - 10166

EP - 10171

JO - PNAS

JF - PNAS

SN - 0027-8424

IS - 27

ER -