Tyrosine phosphorylation of the chlamydial effector protein Tarp is species specific and not required for recruitment of actin

Dawn R Clifton, Cheryl A Dooley, Scott S Grieshaber, Reynaldo A Carabeo, Kenneth A Fields, Ted Hackstadt

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

Chlamydiae are obligate intracellular pathogens that efficiently induce their endocytosis by susceptible eukaryotic host cells. Recently, a Chlamydia trachomatis type III secreted effector protein, Tarp, was found to be translocated and tyrosine phosphorylated at the site of entry and associated with the recruitment of actin that coincides with endocytosis. C. trachomatis Tarp possesses up to six direct repeats of approximately 50 amino acids each. The majority of the tyrosine residues are found within this repeat region. Here we have ectopically expressed distinct domains of Tarp in HeLa 229 cells and demonstrated that tyrosine phosphorylation occurs primarily within the repeat region, while recruitment of actin is mediated by the C-terminal domain of the protein. A comparison of other sequenced chlamydial genomes revealed that each contains an ortholog of Tarp, although Chlamydia muridarum, Chlamydophila caviae, and Chlamydophila pneumoniae Tarp lack the large repeat region. Immunofluorescence and immunoblotting using an antiphosphotyrosine antibody show no evidence of phosphotyrosine at the site of entry of C. muridarum, C. caviae, and C. pneumoniae, although each species similarly recruits actin. Ectopic expression of full-length C. trachomatis and C. caviae Tarp confirmed that both recruit actin but only C. trachomatis Tarp is tyrosine phosphorylated. The data indicate that the C-terminal domain of Tarp is essential for actin recruitment and that tyrosine phosphorylation may not be an absolute requirement for actin recruitment. The results further suggest the potential for additional, unknown signal transduction pathways associated specifically with C. trachomatis.
Original languageEnglish
Pages (from-to)3860-8
Number of pages9
JournalInfection and Immunity
Volume73
Issue number7
DOIs
Publication statusPublished - Jul 2005

Fingerprint

Chlamydia trachomatis
Tyrosine
Actins
Phosphorylation
Chlamydia muridarum
Chlamydophila pneumoniae
Proteins
Endocytosis
Chlamydophila
Phosphotyrosine
Chlamydia
Nucleic Acid Repetitive Sequences
Eukaryotic Cells
HeLa Cells
Immunoblotting
Fluorescent Antibody Technique
Signal Transduction
Genome
Amino Acids
Antibodies

Keywords

  • Repetitive Sequences, Amino Acid
  • Bacterial Proteins
  • Phosphorylation
  • Molecular Sequence Data
  • Actins
  • Tyrosine
  • Amino Acid Sequence
  • Species Specificity
  • Signal Transduction
  • Chlamydia

Cite this

Tyrosine phosphorylation of the chlamydial effector protein Tarp is species specific and not required for recruitment of actin. / Clifton, Dawn R; Dooley, Cheryl A; Grieshaber, Scott S; Carabeo, Reynaldo A; Fields, Kenneth A; Hackstadt, Ted.

In: Infection and Immunity, Vol. 73, No. 7, 07.2005, p. 3860-8.

Research output: Contribution to journalArticle

Clifton, Dawn R ; Dooley, Cheryl A ; Grieshaber, Scott S ; Carabeo, Reynaldo A ; Fields, Kenneth A ; Hackstadt, Ted. / Tyrosine phosphorylation of the chlamydial effector protein Tarp is species specific and not required for recruitment of actin. In: Infection and Immunity. 2005 ; Vol. 73, No. 7. pp. 3860-8.
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AU - Clifton, Dawn R

AU - Dooley, Cheryl A

AU - Grieshaber, Scott S

AU - Carabeo, Reynaldo A

AU - Fields, Kenneth A

AU - Hackstadt, Ted

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N2 - Chlamydiae are obligate intracellular pathogens that efficiently induce their endocytosis by susceptible eukaryotic host cells. Recently, a Chlamydia trachomatis type III secreted effector protein, Tarp, was found to be translocated and tyrosine phosphorylated at the site of entry and associated with the recruitment of actin that coincides with endocytosis. C. trachomatis Tarp possesses up to six direct repeats of approximately 50 amino acids each. The majority of the tyrosine residues are found within this repeat region. Here we have ectopically expressed distinct domains of Tarp in HeLa 229 cells and demonstrated that tyrosine phosphorylation occurs primarily within the repeat region, while recruitment of actin is mediated by the C-terminal domain of the protein. A comparison of other sequenced chlamydial genomes revealed that each contains an ortholog of Tarp, although Chlamydia muridarum, Chlamydophila caviae, and Chlamydophila pneumoniae Tarp lack the large repeat region. Immunofluorescence and immunoblotting using an antiphosphotyrosine antibody show no evidence of phosphotyrosine at the site of entry of C. muridarum, C. caviae, and C. pneumoniae, although each species similarly recruits actin. Ectopic expression of full-length C. trachomatis and C. caviae Tarp confirmed that both recruit actin but only C. trachomatis Tarp is tyrosine phosphorylated. The data indicate that the C-terminal domain of Tarp is essential for actin recruitment and that tyrosine phosphorylation may not be an absolute requirement for actin recruitment. The results further suggest the potential for additional, unknown signal transduction pathways associated specifically with C. trachomatis.

AB - Chlamydiae are obligate intracellular pathogens that efficiently induce their endocytosis by susceptible eukaryotic host cells. Recently, a Chlamydia trachomatis type III secreted effector protein, Tarp, was found to be translocated and tyrosine phosphorylated at the site of entry and associated with the recruitment of actin that coincides with endocytosis. C. trachomatis Tarp possesses up to six direct repeats of approximately 50 amino acids each. The majority of the tyrosine residues are found within this repeat region. Here we have ectopically expressed distinct domains of Tarp in HeLa 229 cells and demonstrated that tyrosine phosphorylation occurs primarily within the repeat region, while recruitment of actin is mediated by the C-terminal domain of the protein. A comparison of other sequenced chlamydial genomes revealed that each contains an ortholog of Tarp, although Chlamydia muridarum, Chlamydophila caviae, and Chlamydophila pneumoniae Tarp lack the large repeat region. Immunofluorescence and immunoblotting using an antiphosphotyrosine antibody show no evidence of phosphotyrosine at the site of entry of C. muridarum, C. caviae, and C. pneumoniae, although each species similarly recruits actin. Ectopic expression of full-length C. trachomatis and C. caviae Tarp confirmed that both recruit actin but only C. trachomatis Tarp is tyrosine phosphorylated. The data indicate that the C-terminal domain of Tarp is essential for actin recruitment and that tyrosine phosphorylation may not be an absolute requirement for actin recruitment. The results further suggest the potential for additional, unknown signal transduction pathways associated specifically with C. trachomatis.

KW - Repetitive Sequences, Amino Acid

KW - Bacterial Proteins

KW - Phosphorylation

KW - Molecular Sequence Data

KW - Actins

KW - Tyrosine

KW - Amino Acid Sequence

KW - Species Specificity

KW - Signal Transduction

KW - Chlamydia

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