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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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DNA elimination analysis of scnRNA production-defective Tetrahymena

ABSTRACT: Analysis of genome-wide IES elimination of Late-scnRNA accumulation-defective cells inducates that Early-scnRNAs are sufficient to induce DNA elimination for a majority of IESs, whereas Late-scnRNAs are important for DNA elimination of some, mainly Type-B, IESs. new MACs of exconjugants were isolated from different mutants at 36 hpm, and the genomic DNA was analyzed by high-throughput sequencing

ORGANISM(S): Tetrahymena thermophila

SUBMITTER: Kazufumi Mochizuki

PROVIDER: E-GEOD-68457 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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DNA elimination analysis of Tetrahymena cells defective in heterochromatin body formation

Project description:In the ciliated protozoan Tetrahymena, de novo heterochromatin body formation is accompanied by programmed DNA elimination. Here, we show that the novel heterochromatin body component Jub1p promotes heterochromatin body formation and dephosphorylation of the Heterochromatin Protein 1 (HP1)-like protein Pdd1p. Through the identification and mutagenesis of the phosphorylated residues of Pdd1p, we demonstrate that Pdd1p dephosphorylation promotes the electrostatic interaction between Pdd1p and RNA in vitro and heterochromatin body formation in vivo. We therefore suggest that heterochromatin bodies are assembled by the Pdd1p-RNA interaction. Jub1p and Pdd1p dephosphorylation are required for heterochromatin body formation and DNA elimination but not for local heterochromatin assembly, indicating that heterochromatin body of itself plays an essential role in DNA elimination. Micronuclei (MICs) and new macronuclei (MACs) of exconjugants were isolated from different mutants at 36 hpm, and the genomic DNA was analyzed by high-throughput sequencing.

2015-12-10 | E-GEOD-70082 | biostudies-arrayexpress

Phosphorylation of an HP1-like protein regulates RNA-bridged heterochromatin body assembly for DNA elimination

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

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Chromatin Immunoprecipitation sequencing of heterochromation associated proteins in Tetrahymena

Project description:In the ciliated protozoan Tetrahymena, de novo heterochromatin body formation is accompanied by programmed DNA elimination. We previously reported that dephosphorylation of the HP1-like protein Pdd1p is required for the formation of heterochromatin bodies during the process of programmed DNA elimination in the ciliated protozoan Tetrahymena. Here, we show that the heterochromatin body component Jub4p is required for Pdd1p phosphorylation, heterochromatin body formation and DNA elimination. Moreover, our analyses of unphosphorylatable Pdd1p mutants demonstrate that Pdd1p phosphorylation is required for heterochromatin body formation and DNA elimination, while it is dispensable for local heterochromatin assembly. Therefore, both phosphorylation and the following dephosphorylation of Pdd1p are necessary to facilitate the formation of heterochromatin bodies. We suggest that Jub4p-mediated phosphorylation of Pdd1p creates a chromatin environment that is a prerequisite for subsequent heterochromatin body assembly and DNA elimination. New macronuclei (MACs) of exconjugants were isolated from wild-type and various mutant cells at 12 hpm (hours post-mixing), sheared chromatin was immunoprecipitated andprecipitated DNA was analyzed by high-throughput sequencing

2016-07-15 | E-GEOD-83249 | biostudies-arrayexpress

Chromatin immunoprecipitation sequencing of heterochromatin-associated proteins in Tetrahymena

Project description:In the ciliated protozoan Tetrahymena, de novo heterochromatin body formation is accompanied by programmed DNA elimination. Here, we show that the novel heterochromatin body component Jub1p promotes heterochromatin body formation and dephosphorylation of the Heterochromatin Protein 1 (HP1)-like protein Pdd1p. Through the identification and mutagenesis of the phosphorylated residues of Pdd1p, we demonstrate that Pdd1p dephosphorylation promotes the electrostatic interaction between Pdd1p and RNA in vitro and heterochromatin body formation in vivo. We therefore suggest that heterochromatin bodies are assembled by the Pdd1p-RNA interaction. Jub1p and Pdd1p dephosphorylation are required for heterochromatin body formation and DNA elimination but not for local heterochromatin assembly, indicating that heterochromatin body of itself plays an essential role in DNA elimination. New macronuclei (MACs) of exconjugants were isolated from wild-type different mutant cells at 12 hpm, shared chromatin was immunoprecipitated and precipitated DNA was analyzed by high-throughput sequencing.

2015-12-10 | E-GEOD-70081 | biostudies-arrayexpress

Jub4p-mediated phosphorylation of Pdd1p is a prerequisite for heterochromatin body assembly and DNA elimination

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2016-07-15 | E-GEOD-83252 | biostudies-arrayexpress

Biased transcription and selective degradation of small RNAs shape the pattern of DNA elimination in Tetrahymena

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2012-06-21 | E-GEOD-38768 | biostudies-arrayexpress

DNA elimination analysis of scnRNA production-defective Tetrahymena

Project description:Analysis of genome-wide IES elimination of Late-scnRNA accumulation-defective cells inducates that Early-scnRNAs are sufficient to induce DNA elimination for a majority of IESs, whereas Late-scnRNAs are important for DNA elimination of some, mainly Type-B, IESs.

2015-06-15 | GSE68457 | GEO

Biased transcription and selective degradation of small RNAs shape the pattern of DNA elimination in Tetrahymena

2012-06-21 | GSE38768 | GEO

Analyses of Early- and Late-scnRNAs of Tetrahymena

Project description:In this study, we demonstrated that there is a novel, unanticipated mechanism regulating programmed DNA elimination: a genome-wide trans-recognition network for IES identification. In this mechanism, Early-scnRNAs produced from Type-A IESs in the MIC identify not only the IESs from which they are derived but also other IESs in trans to trigger the cis-spreading of Late-scnRNA production in the IESs. This cis-spreading of Late-scnRNA production requires heterochromatin formation . Furthermore, these Late-scnRNAs can recognize other IESs in trans. This “chain reaction” of Late-scnRNA production by the trans-recognition network most likely provides strong robustness in DNA elimination by buffering cell-to-cell variability in the initial Early-scnRNA populations.

2015-06-15 | GSE68455 | GEO

Analyses of Early- and Late-scnRNAs of Tetrahymena

2015-06-15 | E-GEOD-68455 | biostudies-arrayexpress

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