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A nuclear export block triggers the decay of newly synthesized polyadenylated RNA

ABSTRACT: A nuclear export block triggers the decay of newly synthesized polyadenylated RNA

PROVIDER: PRJNA427344 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6413660.fastq.gz	Fastqsanger.gz
	SRR6413670.fastq.gz	Fastqsanger.gz
	SRR6413651.fastq.gz	Fastqsanger.gz
	SRR6413661.fastq.gz	Fastqsanger.gz
	SRR6413671.fastq.gz	Fastqsanger.gz

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A nuclear export block triggers the decay of newly synthesized polyadenylated RNA

Project description:Genomes are promiscuously transcribed, necessitating mechanisms that facilitate the sorting of RNA for function or destruction. The polyA (pA) tail is one such distinguishing feature, which in the S. cerevisiae nucleus is bound by the Nab2p protein, yielding transcript protection. As Nab2p also contacts the main nuclear export factor Mex67p, we asked whether transport kinetics contributes to RNA sorting. Indeed, 3’end sequencing of newly transcribed pA+ RNAs demonstrates that nuclear depletion of Mex67p elicits their instant and global decay. A similar phenotype is evident upon inactivation of other export factors and proportional to the amount of nuclear pA+ RNA. Since RNA production is partially rescued by Nab2p over-expression, we propose that an export-block out-titrates Nab2p onto nuclear retained pA+ RNA, reducing the pool of Nab2p free to protect new transcripts. More generally, we suggest that nuclear RNA decay, negotiated by Nab2p availability, aids in balancing cellular transcript supply with demand.

2018-08-28 | GSE108477 | GEO

A Nuclear Export Block Triggers the Decay of Newly Synthesized Polyadenylated RNA.

Project description:Genomes are promiscuously transcribed, necessitating mechanisms that facilitate the sorting of RNA for function or destruction. The polyA (pA) tail is one such distinguishing feature, which in the Saccharomyces cerevisiae nucleus is bound by the Nab2p protein, yielding transcript protection. As Nab2p also contacts the main nuclear export factor Mex67p, we asked whether transport kinetics contributes to RNA sorting. Indeed, 3' end sequencing of newly transcribed pA+ RNAs demonstrates that nuclear depletion of Mex67p elicits their instant and global decay. A similar phenotype is evident upon inactivation of other export factors and proportional to the amount of nuclear pA+ RNA. As RNA expression is partially rescued by Nab2p overexpression, we propose that an export block out-titrates Nab2p onto nuclear-retained pA+ RNA, reducing the pool of Nab2p available to protect new transcripts. More generally, we suggest that nuclear RNA decay, negotiated by Nab2p availability, aids in balancing cellular transcript supply with demand.

| S-EPMC6130047 | biostudies-literature

ALYREF is shared by polyadenylated and nonpolyadenylated mRNA processing and export pathways [iCLIP-seq]

Project description:we try to investigate the binding of ALYREF and NXF1 on histone mRNA when the cell treated with indicated siRNAs. ALYREF plays key roles in nuclear export of polyadenylated mRNAs and also modulates their 3' processing, but whether it is involved in regulating RNAs beyond polyadenylated mRNAs is unknown. The replication-dependent (RD) histone mRNAs are not polyadenylated, but end in a stem-loop (SL) structure. Here we demonstrate that ALYREF prevalently binds a region next to the SL on RD histone mRNAs. SL-binding protein (SLBP) directly interacts with ALYREF and ensures this binding. To examine how SLBP KD impact ALYREF distribution on the histone mRNA, we carried out ALYREF iCLIP in control and SLBP KD cells. To investigate the functional consequence for ALYREF binding on histone mRNAs, we isolated polyA+ and polyA- RNAs from control and ALYREF KD cells, and carried out RNA-seq separately.

2019-02-27 | GSE124735 | GEO

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Transcriptome-based mapping of m6A sites in newly synthesized mRNA population in U2OS cells.

Project description:The goal was to see whether and how mRNA stability is linked to m6A peak position in newly synthesized mRNA population.

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ALYREF is shared by polyadenylated and nonpolyadenylated mRNA processing and export pathways

Project description:ALYREF is shared by polyadenylated and nonpolyadenylated mRNA processing and export pathways

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Newly synthesized proteome analysis of interferon gamma treated Hela cells

Project description:Using a newly-developed workflow for quantitative newly synthesized proteome analysis (QuaNPA), featuring automated sample processing and multiplexed DIA (plexDIA) analysis, changes in the newly synthesized proteome of IFN-gamma treated Hela cells were monitored over time.

2023-10-19 | PXD038915 | Pride

Newly synthesized RNA analysis in WT, SAGA and ATAC mutant mouse embryonic stem cells

Project description:To analyse the genome-wide impact of inactivation of the ATAC or SAGA coactivator complexes on RNA polymerase II (Pol II) transcription, we purified newly synthesized RNA from mutant mouse embryonic stem (ES) cell lines in which subunits of ATAC (Yeats2, Zzz3) or SAGA (Supt7l) are inactivated or depleted. We also performed this analysis in mutant cell lines in which a subunit (Tada3) of the shared histone acetyltransferase activity of these two complexes is depleted. Newly synthesized RNA was purified following the 4sU labelling method (more details in extract protocol). For two wildtype samples, the total RNA input was also analysed.

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Effects of m6A modification on turnover of newly synthesized mRNAs at the transcriptome level.

Project description:We developed a method to deconvolute decays of m6A mRNAs and their unmethylated isoforms from newly synthesized total RNAs across the transcriptome to study the effects of m6A modification on decays of not only the m6A-modified RNA pool but also the unmehnylated isoform pool.

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