Localization and Characterization of GTP-Binding Protein CT703 in the Chlamydia trachomatis-Infected Cells

Current Microbiology - Tập 62 - Trang 465-471 - 2010
Kun Du1, FuYan Wang1, Zhi Huo1, Jie Wang1, Wen Cheng1, Ming Li1, Ping Yu1
1Department of Immunology, Xiangya School of Medicine, Central South University, Changsha, China

Tóm tắt

To localize and characterize the GTP-binding protein encoded by the chlamydial ORF CT703 in the Chlamydia trachomatis-infected cells, the gene coding for CT703 in the Chlamydia trachomatis serovar L2 genome was cloned into the prokaryotic expression vector pGEX and expressed as GST fusion protein in the E. coli BL21 strain. The GST-CT703 fusion protein was purified and used to raise antigen-specific antibodies. Using the anti-fusion protein antibodies, we localized the endogenous CT703 protein inside the chlamydial inclusion using an indirect immunofluorescence assay (IFA). We also detected a significantly decreased level of CT703 in cultures that were induced to undergo persistent infection. These observations suggest that CT703 may be an important regulator for promoting chlamydial productive infection.

Tài liệu tham khảo

Askienazy-Elbharl M, Henry-Suchet J (1999) Persistent “Silent” Chlamydia trachomatis female genital tract infections. Infections Dis Obstet Gynecol 7:31–34 Beatty WL, Byrne GI, Morrison RP (1993) Morphologic and antigenic characterization of interferon γ-mediated persistent Chlamydia trachomatis infection in vitro. Proc Natl Acad Sci USA 90:3998–4002 Beatty WL, Morrison RP et al (1994) Immunoelectron-microscopic quantitation of differential levels of chlamydial proteins in a cells culture model of persistent Chlamydia trachomatis infection. Infect Immun 62:4059–4062 Belland RJ, David E, Virok D et al (2003) Transcription analysis of chlamydial growth IFN-γ-mediated persistence and reactivation. PNAS 100:15971–15976 Carabeo RA, Mead DJ, Hackstadt T (2003) Golgi-dependent transport of cholesterol to the Chlamydia trachomatis inclusion. PNAS 100:6771–6776 Chen CQ, Chen D, Sharma J et al (2006) The hypothetical protein CT813 is localized in the Chlamydia trachomatis inclusion membrane and is immunogenic in women urogenitally infected with C. trachomatis. Infect Immun 74:4826–4840 Fan T, Lu H, Hu H et al (1998) Inhibition of apoptosis in Chlamydia-infected cells: blockade of mitochondrial cytochrome c release and caspase activation. J Exp Med 187:487–496 Hackstadt T (1998) The diverse habitats of obligate intracellular parasites. Curr Opin Microbiol 1:82–87 Hackstadt T, Scidmore MA, DRockey D (1995) Lipid metabolism in Chlamydia trachomatis-infected cells: directed trafficking of Golgi-derived sphingolipids to the chlamydial inclusion. PNAS 92:4877–4881 Hackstadt T, Rockey DD, Heinzen RA et al (1996) Chiamydia trachomatis interrupts an exocytic pathway to acquire endogenously synthesized sphingomyelin in transit from the Golgi apparatus to the plasma membrane. EMBO J 15:964–977 Hackstadt T, Fischer ER, Scidmore MA et al (1997) Origins and functions of the chlamydial inclusion. Trends Microbiol 5:288–293 Hybiske K, Stephens RS (2007) Mechanisms of host cell exit by the intracellular bacterium Chlamydia. Proc Natl Acad Sci USA 104:11430–11435 Kuipers JG, Zeidler H, Kohler L (2003) How does Chlamydia cause arthritis? Rheum Dis Clin North Am 29:613–629 Pinkerton SD, Layde PM, For the NIMH multisite HIV prevention trial group (2002) Using sexually transmitted disease incidence as a surrogate marker for HIV incidence in prevention trials: a modeling study. Sex Transm Dis 29:298–307 Polkinghorn A, Hogan RJ, Vaughan L et al (2006) Differential expression of chlamydial signal transduction genes in normal and interferon gamma-induced persistent Chlamydophila pneumoniae infections. Microbes Infect 8:61–72 Rzomp KA, Scholtes LD, Briggs BJ et al (2003) Rab GTPases are recruited to Chlamydial inclusions in both a species-dependent and species-independent manner. Infect Immun 71:5855–5870 Sambrook J, Ruessell DW (2002) Molecular cloning: a laboratory manual, 3rd edn. Science Press, Beijing, pp 1217–1265 Scidmore MA, Fischer ER, Hackstadt T (1996) Sphingolipids and glycoproteins are differentially trafficked to the Chlamydia trachomatis inclusion. J Cell Biol 134:363–374 Scidmore MA, Fischer ER, Hackstadt T (2003) Restricted fusion of Chlamydia trachomatis vesicles with endocytic compartments during the initial stages of infection. Infect Immun 71:973–984 Sherman KJ, Daling JR, Stergachis A et al (1990) Sexually transmitted diseases and tubal pregnancy. Sex Transm Dis 17:115–121 Stephens RS, Kalman S, Lammel C et al (1998) Genome sequence of an obligate intracellular pathogen of humans: Chlamydia trachomatis. Science 282:754–759 Su H, McClarty G, Dong F et al (2004) Activation of Raf/MEK/ERK/cPLA2 signaling pathway is essential for Chlamydia acquisition of host glycerophomatis. J Biol Chem 279:9409–9416 Taylor HR, Johnson SL, Schachter J et al (1987) Pathogenesis of trachoma: the stimulus for inflammation. J Immunol 138:3023–3027 Wallin KL, Wiklund F, Luostarinen T et al (2002) A population-based prospective study of Chlamydia trachomatis infection and cervical carcinoma. Int J Cancer 101:371–374 Xiao Y, Zhong Y, Greene W et al (2004) Chlamydia trachomatis infection inhibits both Bax and Bak activation induced by staurosporine. Infect Immun 72:5470–5474 Zhong G, Fan P, Ji H et al (2001) Identification of a Chlamydial protease-like activity factor responsible for the degradation of host transcription factor. J Exp Med 193:935–942