Ontology highlight
ABSTRACT:
PROVIDER: PRJNA395910 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR5871001.fastq.gz | Fastqsanger.gz |
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Project description:Gigantopithecus blacki was a giant hominid that inhabited Southeast Asia during the Pleistocene. Its evolutionary relationship to other great ape species, and their divergence during the Middle and Late Miocene (16-5.3 Mya), remains disputed. In part, this is due to the absence of cranial and postcranial remains and size-induced allometry. Proposed hypothesis on the phylogenetic positions of Gigantopithecus have therefore been wide-ranging among hominoids, but none has received independent validation based on molecular evidence. To clarify the phylogenetic placement of Gigantopithecus blacki, we retrieved enamel proteome sequences from a 1.9 million years (Mya) old molar found in Chuifeng Cave, China. We demonstrate that Gigantopithecus is most closely related to orangutans (genus Pongo). We also estimate the Gigantopithecus-Pongo divergence to about 10-12 Mya, implying its speciation is part of the Miocene radiation of great apes. These sequences are approximately 6 times older, in a normalized thermal context, than any previously published mammalian proteome or genome. The survival of an Early Pleistocene dental enamel proteome in the subtropics further expands the scope of palaeoproteomic analysis into geographic areas and time periods previously considered incompatible with biomolecular preservation.
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Project description:Administration of attenuated autologous Theileria parva infected cells can be used as an alternative to the infection-and-treatment method for inducing immunological protection against East Coast Fever. The mechanism of attenuation however has not been described. Using RNA sequencing, the transcriptomes of both host and parasite in uninfected (control), pathogenic (day 7 post-infection) and attenuated (day 69 post-infection) T. parva infected bovine CD4+ T-cells were characterized and compared. Our findings suggest that three major mechanisms are associated with attenuation of T. parva-infected cells – a decrease in proliferation, a partial restoration of the inflammatory profile, and a shift in metabolism. Several host genes (TRAIL, PD-1, TGF-β and granzymes) were identified as candidates for further exploration. Evaluation of the parasite transcriptomes in these cells also provided first insights into potential candidate T. parva genes involved in attenuation, but subsequent studies are required to further examine these.
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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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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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