Project description:In order to resolve controversies concerning SMYD's substrates and functions, we studied SMYD1 (TTHERM_00578660), the only homologue in the unicellular eukaryote Tetrahymena thermophila. We epitope-tagged SMYD1, and analyzed its localization and interactome. We also characterized ΔSMYD1 cells, focusing on the replication and transcription phenotype. Our results show that: 1) SMYD1 is present in both cytoplasm and transcriptionally active macronucleus and shuttles between cytoplasm and macronucleus, which is consistent with its association with both histone and non-histone substrates; 2) SMYD1 is involved in DNA replication and regulates transcription of metabolism-related genes; 3) HSP90 is an potential substrate for SMYD1 and it may regulate target selection of HSP90, leading to pleiotropic effects in both the cytoplasm and the nucleus.
Project description:We present a comprehensive transcriptome of ciliate T. thermophila using the Illumina RNA-seq platform. The data was generated from the six mRNA samples of growth, starvation and conjugation of Tetrahymena. Despite an AT rich genome, there are about 124.6 million reads mapped to T. thermophila genome. Using these mapped reads, we have significantly improved the previous genome annotation and investigated the gene expression. Besides, our result also provided a comprehensive understanding of the alternative splicing in T. thermophila, and suggested the existence of the regulated unproductive splicing and translation (RUST) in the single-celled eukaryote. RNA-seq for six samples of Tetrahymena growth, starvation and conjugation.
Project description:We present a comprehensive transcriptome of ciliate T. thermophila using the Illumina RNA-seq platform. The data was generated from the six mRNA samples of growth, starvation and conjugation of Tetrahymena. Despite an AT rich genome, there are about 124.6 million reads mapped to T. thermophila genome. Using these mapped reads, we have significantly improved the previous genome annotation and investigated the gene expression. Besides, our result also provided a comprehensive understanding of the alternative splicing in T. thermophila, and suggested the existence of the regulated unproductive splicing and translation (RUST) in the single-celled eukaryote.
Project description:To identified the differential expressed genes between the arsenic exposure cells and wild type cells As is a well- known carcinogen, it receives special attention. Due to its ubiquitous distribution in the environment, a unicellular model system related to the environment will be useful in toxicological research of arsenic. The eukaryotic protist Tetrahymena thermophila is a well-established model in classical toxicology and now a potential model at genomic level. In the present study, Tetrahymena thermophila was inbred with arsenate. With the use of microarray technique, the genes differentially expressed at 4 time points in exposure to arsenate were screened out. The major biological processes involved and their changes over time were identified with enrichment analysis and transformed expression data. Characteristics of arsenic toxicity and response to arsenic were identified, proving T. thermophila a good unicellular model for arsenic toxicological research at genomic level.
Project description:As a prototypic ciliated protozoan, Tetrahymena thermophila harbors two functionally and physically distinct nuclei within a shared cytoplasm. During the vegetative phase of the life cycle, the S phases of the diploid germline micronucleus and polyploid somatic macronucleus are temporally offset. Here we report the first RNA-Seq analysis across the cell cycle of a binucleated ciliated protozoan. We then performed gene expression profiling analysis using data obtained from RNA-seq of 4 different cells at two time points.
Project description:In endolysosomal networks, two hetero-hexameric tethers called HOPS and CORVET are found widely throughout eukaryotes. The ciliate Tetrahymena thermophila is a unicellular organism with elaborate endolysosomal pathways. Curiously, Tetrahymena and related protozoa have lost HOPS. Tetrahymena encodes multiple paralogs of most CORVET subunits, assembled into six distinct hexameric complexes, including a distinct Vps8 subunit. Here we analyze the composition of these complexes immunoisolated by affinity capture using the Vps8-FLAG paralogs as bait.
Project description:Microarray analyses were performed to compare the gene expression profiles of wild-type and several mutant strains of the ciliated protozoan Tetrahymena thermophila. Elimination of H3K4 methylation (hht2-K4Q) and knockout of either of the ubiquitylation enzymes (delta-UBC2 and delta-BRE1) affects a broader spectrum of genes than elimination of H2B ubiquitylation (htb1-K115R).