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Investigation of the RNMT/RAM RNA interactome in mouse embryonic stem cells [Brain_seCLIP]

ABSTRACT: Investigation of the RNMT/RAM RNA interactome in mouse embryonic stem cells [Brain_seCLIP]

PROVIDER: PRJNA816065 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR18324500_1.fastq.gz	Fastqsanger.gz
	SRR18324500_2.fastq.gz	Fastqsanger.gz
	SRR18324501_1.fastq.gz	Fastqsanger.gz
	SRR18324501_2.fastq.gz	Fastqsanger.gz
	SRR18324502_1.fastq.gz	Fastqsanger.gz

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Investigation of the RNMT/RAM RNA interactome in mouse embryonic stem cells

Project description:Pilot investigation to determine the endogenouse RNA-binding targets of the cap methlytransferase complex RNMT/RAM in mouse ES cells

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Investigation of the RNMT/RAM RNA interactome in mouse embryonic stem cells [Brain_seCLIP]

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mRNA cap methyltransferase, RNMT-RAM, promotes RNA pol II transcription

Project description:mRNA cap addition occurs early during RNA pol II transcription, facilitating pre-mRNA processing and translation. We report that the mammalian mRNA cap methyltransferase, RNMT-RAM, promotes RNA pol II transcription, independently of mRNA capping and translation. In cells, sub-lethal suppression of RNMT-RAM reduces RNA pol II occupancy, net mRNA synthesis and pre-mRNA levels. Conversely, expression of RNMT-RAM increases transcription independently of cap methyltransferase activity. In isolated nuclei, recombinant RNMT-RAM stimulates transcriptional output; this requires the RAM RNA-binding domain. RNMT-RAM interacts with nascent transcripts along their entire length and with transcription associated factors including RNA pol II subunits, SPT4, SPT6 and PAFc. Suppression of RNMT-RAM inhibits transcriptional markers including histone H2B K120 ubiquitination, H3 K4 and K36 methylation, RNA pol II S5 and S2 phosphorylation and PAFc recruitment. These findings suggest that multiple interactions between RNMT-RAM, RNA pol II factors and RNA along the transcription unit stimulate transcription.

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Rnmt and Ram regulate the activated CD4 T cell transcriptome

Project description:We investigated the role of RNMT and its cofactor RAM in T cells using conditional knockout mouse models. We demonstrate that RNMT and RAM have broadly overlapping roles in CD4 T cell activation.

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Project description:Rnmt and Ram regulate the activated CD4 T cell transcriptome

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RNMT regulates the naïve CD4 T cell transcriptome

Project description:We investigated the role of RNMT in T cells using an Rnmt conditional knockout mouse model. We report that the mRNA cap methyltransferase, RNMT, supports naïve T cell survival and activation-induced proliferation. We demonstrate that RNMT has gene-specific impacts in T cells, selectively regulating expression of terminal polypyrimidine tract (TOP) mRNAs which are targets of the m7G-cap binding protein LARP1. These RNAseq experiments investigate the effect of Rnmt cKO on the naïve CD4 T cell transcriptome.

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Psuedouridine sequencing rRNA in Rnmt cKO CD4 T cells

Project description:We investigated the role of RNMT in T cells using an Rnmt conditional knockout mouse model. We report that the mRNA cap methyltransferase, RNMT, supports naïve T cell survival and activation-induced proliferation. We demonstrate that RNMT has gene-specific impacts in T cells, selectively regulating expression of terminal polypyrimidine tract (TOP) mRNAs and small RNAs including snoRNAs. To deterine if had any effect on ribosomal RNA modification we carried out pseudoridine sequencing.

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