Transforming Growth Factor α Binds to
            <i>Trypanosoma cruzi</i>
            Amastigotes To Induce Signaling and Cellular Proliferation

Alexander, A. D.; Villalta, Fernando; Lima, Maria F.

doi:10.1128/iai.71.7.4201-4205.2003

Cited by 7 publications

(7 citation statements)

References 22 publications

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“…Cell monolayers were solubilized in 0.1% CHAPS containing protease inhibitors [13]. The same protein concentrations of all samples were separated by SDS–PAGE, transferred onto nitrocellulose membranes, probed with goat IgG to TSP‐1, anti‐fibronectin, anti‐laminin, or anti‐galectin‐3 antibodies, developed using the Western Breeze chemiluminescence kit (Invitrogen) as described [14]. Blots were stripped and reprobed with a monoclonal IgG to β‐actin.…”

Section: Methodsmentioning

confidence: 99%

Stable RNA interference of host thrombospondin‐1 blocks Trypanosoma cruzi infection

et al. 2006

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Interactions between Trypanosoma cruzi and the extracellular matrix play an important role in cellular invasion. Here we show that T. cruzi increases the levels of thrombospondin-1 (TSP-1) expression in host cells during early infection. Stable RNA interference of host cell TSP-1 knocks down the levels of TSP-1 transcripts and protein expression in mammalian cells causing inhibition of T. cruzi infection. Addition of TSP-1 to these cells restores infection. Thus, host TSP-1, regulated by the parasite, plays a crucial role in early infection. This is the first report showing that a human parasite modulates TSP-1 expression to facilitate infection.

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Section: Methodsmentioning

confidence: 99%

Stable RNA interference of host thrombospondin‐1 blocks Trypanosoma cruzi infection

et al. 2006

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“…Figure 3A (left panel) shows that antibodies to human galectin-3 immunoprecipitate biotinylated surface galectin-3, whereas preimmune antibodies do not. To investigate whether galectin-3 is released into the cellular medium, we analyzed immunoblots of supernatant cultures of CASM cells incubated in Dulbecco's minimum essential medium (DMEM) for 6 h or in DMEM alone as described previously (2,26,28). Figure 3A (right panel) shows that antibodies to galectin-3 detect galectin-3 on the culture supernatant of CASM cells, whereas anti-galectin-3 antibodies did not react with medium alone, indicating that galectin-3 is released by CASM cells into the culture supernatant.…”

mentioning

confidence: 99%

Human Galectin-3 PromotesTrypanosoma cruziAdhesion to Human Coronary Artery Smooth Muscle Cells

Kleshchenko¹,

Moody²,

Furtak

et al. 2004

Infect Immun

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“…Trypomastigotes, epimastigotes, and amastigotes (2 ϫ 10 7 ) were solubilized in 0.1% CHAPS {3-[(3-cholamidylpropyl)-dimethylammonio]-1-propanesulfonate} containing protease inhibitors (GE Healthcare, Piscataway, NJ), and the same protein concentrations were loaded onto a 12% SDS-PAGE gel, transferred onto nitrocellulose membranes, probed with goat anti-Tc-1 IgG, and developed with peroxidase-conjugated mouse anti-goat IgG by ECL (GE Healthcare, Piscataway, NJ) as described previously (2). Blots were stripped and reprobed with monoclonal IgG specific for ␤-actin.…”

Section: Cruzi Ckii Substrate Mediates Cellular Infection 3923mentioning

confidence: 99%

“…The same volume of immunoprecipitates was separated by SDS-PAGE, blotted onto nitrocellulose membranes, and probed with IgG specific for Tc-1. Blots were incubated with avidin-horseradish peroxidase and developed by ECL (GE Healthcare) as described previously (2).…”

Section: Cruzi Ckii Substrate Mediates Cellular Infection 3923mentioning

confidence: 99%

Molecular Cloning of a Trypanosoma cruzi Cell Surface Casein Kinase II Substrate, Tc-1, Involved in Cellular Infection

et al. 2006

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In this work, we report the cloning and characterization of the first cell surface casein kinase II (CKII) substrate (Tc-1) of Trypanosoma cruzi, the causative agent of Chagas' disease. Analysis of the gene sequence revealed a 1,653-bp open reading frame coding for 550 amino acid residues. Northern blot analysis showed a 4.5-kb transcript that is expressed in invasive trypomastigotes but not in noninvasive epimastigote forms of T. cruzi. Southern blot analysis indicates that Tc-1 is a single-copy gene. At the amino acid level, Tc-1 displayed 95% and 99% identity to two hypothetical proteins recently reported by the T. cruzi genome project. Analysis of the translated amino acid sequence indicates that the Tc-1 gene has a putative transmembrane domain with multiple cytoplasmic and extracellular CKII phosphosites. Exogenous human CKII was able to phosphorylate serine residues on both recombinant Tc-1 and Tc-1 of intact trypomastigotes. This phosphorylation was inhibited by the CKII inhibitors heparin and 4,5,6,7,-tetrabromo-2-azabenzimidazole. Immunoblots of solubilized trypomastigotes, epimastigotes, and amastigotes probed with anti-recombinant Tc-1 immunoglobulin G revealed a 62-kDa protein that is expressed only in infective trypomastigotes. Immunoprecipitation of labeled surface proteins of trypomastigotes indicated that the 62-kDa protein is a surface protein, and we found that the protein is uniformly distributed on the surface of trypomastigotes by direct immunofluorescence. Antibodies to Tc-1 effectively blocked trypomastigote invasion of host cells and consequently reduced parasite load. Preincubation of either trypomastigotes or myoblasts with CKII inhibitors blocked T. cruzi infection. Thus, for the first time, we describe a cell surface CKII substrate of a protozoan parasite that is phosphorylated by human CKII and that is involved in cellular infection.

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Transforming Growth Factor α Binds to Trypanosoma cruzi Amastigotes To Induce Signaling and Cellular Proliferation

Cited by 7 publications

References 22 publications

Stable RNA interference of host thrombospondin‐1 blocks Trypanosoma cruzi infection

Stable RNA interference of host thrombospondin‐1 blocks Trypanosoma cruzi infection

Human Galectin-3 PromotesTrypanosoma cruziAdhesion to Human Coronary Artery Smooth Muscle Cells

Molecular Cloning of a Trypanosoma cruzi Cell Surface Casein Kinase II Substrate, Tc-1, Involved in Cellular Infection

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