Characterization of a Secretory Type Theileria parva Glutaredoxin Homologue Identified by Novel Screening Procedure

Ebel, Thomas; Middleton, J.; Frisch, Angelika; Lipp, Joachim

doi:10.1074/jbc.272.5.3042

Cited by 23 publications

(11 citation statements)

References 49 publications

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“…Conserved proteins on the outer membrane surface of the parasite or secreted proteins represent ideal candidates for parasite host cell interactions (ole-MoiYoi 1995). Until now, some Theileria proteins have been described which are inserted into the schizont's membrane (Baylis et al 1993;Ebel et al 1997;Skilton et al 1998;Schnittger et al 2002;Schneider et al 2004). However, there are few investigations on molecules potentially interfering with signaling pathways of the host.…”

Section: Introductionmentioning

confidence: 99%

Schizonts of Theileria annulata interact with the microtubuli network of their host cell via the membrane protein TaSP

et al. 2010

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Intracellular leukoproliferative Theileria are unique as eukaryotic organisms that transform the immune cells of their ruminant host. Theileria utilize the uncontrolled proliferation for rapid multiplication and distribution into host daughter cells. The parasite distribution into the daughter cells is accompanied by a tight association with the host cell mitotic apparatus. Since the molecular basis for this interaction is largely unknown, we investigated the possible involvement of the immunodominant Theileria annulata surface protein, TaSP, in the attachment of the parasite to host cell microtubule network. Confocal microscopic analyses showed co-localization of the TaSP protein with alpha-tubulin and reciprocal immuno-co-precipitation experiments demonstrated an association of TaSP with alpha-tubulin in vivo. In addition, the partially expressed predicted extracellular domain of TaSP co-localized with the mitotic spindle of dividing cells and was co-immunoprecipitated with alpha-tubulin in transiently transfected Cos-7 cells devoid of other T. annulata expressed proteins. Pull-down studies showed that there is a direct interaction between TaSP and polymerized microtubules. Analysis of the interaction of TaSP and host microtubulin during host cell mitosis indicated that TaSP co-localizes and interacts with the spindle poles, the mitotic spindle apparatus and the mid-body. Moreover, TaSP was demonstrated to be localized to the microtubule organizing center and to physically interact with gamma-tubulin. These data support the notion that the TaSP-microtubule interaction may be playing a potential role in parasite distribution into daughter host cells and give rise to the speculation that TaSP may be involved in regulation of microtubule assembly in the host cell.

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Section: Introductionmentioning

confidence: 99%

Schizonts of Theileria annulata interact with the microtubuli network of their host cell via the membrane protein TaSP

et al. 2010

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“…Conserved proteins on the outer membrane surface of the parasite or secreted proteins represent ideal candidates for parasite-host cell interactions. Until now, some Theileria proteins have been described, which are inserted into the schizont's membrane (Baylis et al 1993;Ebel et al 1997;Skilton et al 1998;Schnittger et al 2002). Members of the TashAT gene family have been postulated to be secreted and transported to the host cell nucleus (Swan et al 1999(Swan et al , 2003Shiels et al 2004).…”

Section: Introductionmentioning

confidence: 99%

Identification, molecular characterization and subcellular localization of a Theileria annulata parasite protein secreted into the host cell cytoplasm

et al. 2007

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Intracellular leucoproliferative Theileria are unique as eukaryotic organisms that transform the immune cells of their ruminant host. Theileria utilize the uncontrolled proliferation for rapid multiplication and distribution into host daughter cells. The equal distribution of the schizont into the daughter cells is thought to be accomplished by a tight association with the host cell mitotic apparatus. In this study, we describe a highly conserved novel 37 kD Theileria annulata protein (TaSE). TaSE was found to be localized inside the parasite, the parasite membrane and within the host cell cytoplasm. Moreover, it co-localized at distinct points with host cell microtubules, which was especially apparent during mitosis, where co-localization was found with the centromere, the mitotic spindle and the midbody. Association of TaSE with the host cell tubulin network was corroborated by coimmunoprecipitation and transient transfection experiments. This is the first description of a theilerial protein co-localizing and potentially interacting with a host cell protein. The distribution of TaSE during mitosis makes it a protein to consider as playing a potential role for parasite distribution into daughter host cells.

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“…In order to identify novel secreted and transmembrane gene products of P. falciparum involved in host-parasite interactions, a P. falciparum cDNA library was screened in an in vitro transcription-translation-translocation assay for genes encoding protein products that are translocated into microsomes and are therefore protected from proteinase K digestion (6). One of the plasmids, designated D13, gave rise to a 37-kDa protein product that resisted proteinase K digestion indicative for membrane association or translocation (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…A total membrane fraction was collected by ultracentrifugation at 100,000 ϫ g for 1 h at 4°C. For certain experiments, equivalent amounts of the sediments were treated with 100 mM Na 2 CO 3 , pH 11.3, or with distilled H 2 O containing protease inhibitors for 30 min on ice prior to further centrifugation for 1 h at 100,000 ϫ g (6). Samples were solubilized in denaturing SDS sample buffer and equivalent amounts of each sample were subjected to Western blot analysis.…”

Section: Identification Of Orf D13mentioning

confidence: 99%

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Identification and Characterization of a Conserved, Stage-Specific Gene Product of Plasmodium falciparum Recognized by Parasite Growth Inhibitory Antibodies

et al. 2003

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We have identified a novel conserved protein of Plasmodium falciparum, designated D13, that is stagespecifically expressed in asexual blood stages of the parasite. The predicted open reading frame (ORF) D13 contains 863 amino acids with a calculated molecular mass of 99.7 kDa and displays a repeat region composed of pentapeptide motives. Northern blot analysis with lysates of synchronized blood stage parasites showed that D13 is highly expressed at the mRNA level during schizogony. The first N-terminal 138 amino acids of D13 were expressed in Escherichia coli and the purified protein was used to generate anti-D13 monoclonal antibodies (MAbs). Using total lysates of blood stage parasites and Western blot analysis, these MAbs stained one single band of ϳ100 kDa, corresponding to the predicted molecular mass of ORF D13. Western blot analysis demonstrated further that D13 is expressed during schizogony, declines rapidly in early ring stages and is undetectable in trophozoites. D13 protein is localized in individual merozoites in a distinct area, as demonstrated by indirect immunofluorescence analysis. After subcellular fractionation, D13 was confined to the pelleted fraction of the parasite lysate and its extraction by alkaline carbonate buffer treatment indicated that D13 is not a membrane-integral protein. Inclusion of certain anti-D13 MAbs into in vitro cultures of blood stage parasites resulted in considerable reduction in parasite growth. The N-terminal domain encompassing 158 amino acids is 94 and 95%, respectively, identical at the amino acid level between Plasmodium knowlesi, Plasmodium yoelii, and P. falciparum. In summary, we describe a novel stage-specifically expressed, highly conserved gene product of P. falciparum that is recognized by parasite growth inhibitory antibodies.

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Characterization of a Secretory Type Theileria parva Glutaredoxin Homologue Identified by Novel Screening Procedure

Cited by 23 publications

References 49 publications

Schizonts of Theileria annulata interact with the microtubuli network of their host cell via the membrane protein TaSP

Schizonts of Theileria annulata interact with the microtubuli network of their host cell via the membrane protein TaSP

Identification, molecular characterization and subcellular localization of a Theileria annulata parasite protein secreted into the host cell cytoplasm

Identification and Characterization of a Conserved, Stage-Specific Gene Product of Plasmodium falciparum Recognized by Parasite Growth Inhibitory Antibodies

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