Theileria parva strain Muguga
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA16138 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| AAGK01.dat.gz | Other | |||
| AAGK01.fasta.gz | Fasta.gz | |||
| AAGK01.master.dat | Other | |||
| SRR3001169_1.fastq.gz | Fastqsanger.gz | |||
| SRR3001169_2.fastq.gz | Fastqsanger.gz |
Items per page: 5 1 - 5 of 6 |
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Project description:Intracellular pathogens develop elaborate mechanisms to survive within the hostile environments of host cells. Theileria parasites infect bovine leukocytes and cause devastating diseases in cattle in developing countries. Theileria spp. have evolved sophisticated strategies to hijack host leukocytes, inducing proliferative and invasive phenotypes characteristic of cell transformation. Intracellular Theileria parasites secrete proteins into the host cell and recruit host proteins to induce oncogenic signaling for parasite survival. It is unknown how Theileria parasites evade host cell defense mechanisms, such as autophagy, to survive within host cells. Here, we show that Theileria annulata parasites sequester the host eIF5A protein to their surface to escape elimination by autophagic processes. We identified a small-molecule compound that reduces parasite load by inducing autophagic flux in host leukocytes, thereby uncoupling Theileria parasite survival from host cell survival. We took a chemical genetics approach to show that this compound induced host autophagy mechanisms and the formation of autophagic structures via AMPK activation and the release of the host protein eIF5A which is sequestered at the parasite surface. The sequestration of host eIF5A to the parasite surface offers a strategy to escape elimination by autophagic mechanisms. These results show how intracellular pathogens can avoid host defense mechanisms and identify a new anti-Theileria drug that induces autophagy to target parasite removal.
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Project description:The protozoan parasite Theileria parva infects and transforms bovine lymphocytes inducing uncontrolled proliferation. The transforming schizont resides free in the host cell cytoplasm and it is assumed that proteins released from the parasite contribute to host cell transformation and parasite persistence. The identification and characterisation of parasite genes encoding candidate secreted proteins constitutes a first step towards elucidating this complex process. In earlier work, it was shown that the genes encoding subtelomere-encoded variable secreted proteins (SVSPs) are located at the subtelomeres of all four T. parva chromosomes and, with 85 members, form the largest Theileria gene family. The majority of predicted proteins contain signal peptides, suggesting secretion into the host cell cytoplasm. We analysed SVSP expression in T. parva-transformed cell lines established in vitro by infection of T or B lymphocytes with cloned T. parva parasites. Preliminary microarray followed by quantitative real-time PCR analysis revealed mRNA expression for a wide range of SVSP genes. The pattern of mRNA expression was neither influenced by the cell type transformed by T. parva, nor by the animal background. Instead, the pattern of SVSP mRNA expression was largely defined by the parasite genotype and found to be relatively stable when monitored in vitro over a period of two months. Experiments using antibodies raised against the SVSP TP03_0882 provided first evidence for protein expression. Interestingly, results indicate that SVSP expression in cell lines established from a cloned parasite is limited to only a small percentage of parasites, suggesting SVSP expression by individual parasites is restricted. Expression of epitope-tagged TP03_0882 in mammalian cells revealed nuclear translocation and localisation to different nuclear compartments, including nucleoli, nucleoplasm and other nuclear bodies. Nuclear translocation to the mammalian cell nucleus was shown to involve two different types of nuclear localisation signals present in the conserved C-terminal region of TP03_0882. This first characterisation, opens up possibilities for future studies on the regulation of gene expression and the biological role of these enigmatic proteins. Expression patterns of SVSP genes in different T. parva-infected cloned cell lines Keywords: Gene expression
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Project description:The protozoan parasite Theileria parva infects and transforms bovine lymphocytes inducing uncontrolled proliferation. The transforming schizont resides free in the host cell cytoplasm and it is assumed that proteins released from the parasite contribute to host cell transformation and parasite persistence. The identification and characterisation of parasite genes encoding candidate secreted proteins constitutes a first step towards elucidating this complex process. In earlier work, it was shown that the genes encoding subtelomere-encoded variable secreted proteins (SVSPs) are located at the subtelomeres of all four T. parva chromosomes and, with 85 members, form the largest Theileria gene family. The majority of predicted proteins contain signal peptides, suggesting secretion into the host cell cytoplasm. We analysed SVSP expression in T. parva-transformed cell lines established in vitro by infection of T or B lymphocytes with cloned T. parva parasites. Preliminary microarray followed by quantitative real-time PCR analysis revealed mRNA expression for a wide range of SVSP genes. The pattern of mRNA expression was neither influenced by the cell type transformed by T. parva, nor by the animal background. Instead, the pattern of SVSP mRNA expression was largely defined by the parasite genotype and found to be relatively stable when monitored in vitro over a period of two months. Experiments using antibodies raised against the SVSP TP03_0882 provided first evidence for protein expression. Interestingly, results indicate that SVSP expression in cell lines established from a cloned parasite is limited to only a small percentage of parasites, suggesting SVSP expression by individual parasites is restricted. Expression of epitope-tagged TP03_0882 in mammalian cells revealed nuclear translocation and localisation to different nuclear compartments, including nucleoli, nucleoplasm and other nuclear bodies. Nuclear translocation to the mammalian cell nucleus was shown to involve two different types of nuclear localisation signals present in the conserved C-terminal region of TP03_0882. This first characterisation, opens up possibilities for future studies on the regulation of gene expression and the biological role of these enigmatic proteins. Expression patterns of SVSP genes in different T. parva-infected cloned cell lines Keywords: Gene expression We performed a three-condition experiment comparing SVSP gene transcription in three T. parva infected cell lines: (1) 211T-A3 (consisting of CD4+ T cells), (2) 211B-A3 (consisting of B cells), and 951T-F44 (consisting of CD4+ T cells). We used direct two-color experiment design, comparing each cell line with the other, i.e. (1) 211T-A3 vs 211B-A3, (2) 211T-A3 vs 951T-F44 and (3) 211B-A3 vs 951T-F44. There were two biological replicates for each hybridization and dye-flips for each hybridization pair. Microarray consisted of four in-slide replicates peer gene as well as no-DNA spots as negative controls.
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