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Small RNA and total RNA sequencing of C. elegans with mutations in the hrde-1 and emb-4 genes

ABSTRACT: Small RNA and total RNA sequencing of C. elegans with mutations in the hrde-1 and emb-4 genes

PROVIDER: PRJEB14792 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR1530690.fastq.gz	Fastqsanger.gz
	ERR1530700.fastq.gz	Fastqsanger.gz
	ERR1530710.fastq.gz	Fastqsanger.gz
	ERR1530720.fastq.gz	Fastqsanger.gz
	ERR1530691.fastq.gz	Fastqsanger.gz

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The activity of the Caenorhabditis elegans Aquarius helicase EMB-4 enables the nuclear RNAi machinery to engage nascent RNA

Project description:Germline small RNA pathways initiate silencing of repetitive elements in animals and an interplay of nuclear small RNAs and chromatin modifications maintain this silencing, protecting the germline from spreading of transposable elements. In C. elegans germline, nuclear argonaute protein HRDE-1 initiates the transcriptional silencing pathway that is crucial for long term and heritable silencing of genes and repetitive regions. Here, we show that HRDE-1 interacts with components of the splicing machinery and the exon-junction complex. One such factor is the conserved RNA helicase EMB-4/AQR that binds introns and recruits the exon-junction proteins to newly spliced RNA. Our data shows that EMB-4/AQR is required for the transcriptional silencing pathway initiated by HRDE-1 and it functions by removing the intronic barriers to silencing thorugh its helicase function.

2017-06-30 | PXD004416 | Pride

The Conserved Intron Binding Protein EMB-4 Plays Differential Roles in Germline Small RNA Pathways of C. elegans

Project description:Proper regulation of the germline transcriptome is essential for the maintenance of fertility and survival of a species. In C. elegans, germline transcriptome homeostasis hinges on a complex repertoire of small RNA pathways that act in both activating and silencing capacities. Our understanding of how fundamental RNA processing steps intersect with these small RNA machineries in the germline remains relatively limited. Here, we link the conserved intron binding protein and splicing factor, EMB-4/AQR/IBP160 to two key 22G-RNA pathways in the C. elegans germline. EMB-4 associates with the Argonautes CSR-1 and HRDE-1, and is enriched at the genomic loci of CSR-1 and HRDE-1 target genes. Loss of emb-4 leads to distinct alterations in CSR-1 vs. HRDE-1 small RNA and mRNA transcriptomes. Our transcriptome-wide analysis shows that EMB-4 is enriched along pre-mRNAs of nearly 10,000 transcripts. For a subset of these genes, including mostly CSR-1 pathway targets, EMB-4 enriches for intronic, but not exonic, sequences. Together these data point to EMB-4 as a factor that may help to distinguish the targets of these two germline small RNA pathways.

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Small RNA and total RNA sequencing of C. elegans with mutations in the hrde-1 and emb-4 genes

Project description:Synchronised animals were grown to young adult stage at 20C on HB101 seeded NGM plates. Animals were harvested and washed 3X in M9 buffer. Settled animals were mixed with Trisure reagent, bead beaten using zirconia beads, and the RNA is extracted using chloroform. For total RNA sequencing, Illumina Ribozero kit is used to remove ribosomal RNA from 1ug of total RNA prior to library preparation. RNA sequencing libraries are prepared using NEB Next Ultra library preparation kit. Small RNA sequencing is performed by treating 5ug of total RNA with Epicentre 5 polyphosphatase to remove the 5 triphosphate. After treatment, RNA is purified by phenol/chloroform extraction and 1ug of RNA is used to prepare small RNA libraries using Illumina TruSeq small RNA library preparation kit. Ribosomal depleted RNA and small RNA libraries are sequenced using Illumina HiSeq 1500 platform.

2017-07-01 | E-MTAB-4877 | biostudies-arrayexpress

Histone H3K9me3 ChIP-seq of young adult C. elegans wildtype, and mutants emb-4 (qm31) and hrde-1 (tm1200)

Project description:Histone H3K9me3 ChIP-seq of young adult C. elegans wildtype, and mutants emb-4 (qm31) and hrde-1 (tm1200)

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Germ granule association drives small RNA specificity for a nuclear Argonaute protein

Project description:RNA interference (RNAi) is a conserved gene silencing process that exists in diverse organisms to protect genome integrity and regulate gene expression. In C. elegans, the majority of RNAi pathway proteins localize to perinuclear, phase-separated germ granules, which are comprised of sub-domains referred to as P granules, Mutator foci, Z granules, and SIMR foci. However, the protein components and function of the newly discovered SIMR foci are unknown. Here we demonstrate that HRDE-2 localizes to SIMR foci and interacts with the germline nuclear RNAi Argonaute HRDE-1. Furthermore, HRDE-1 also localizes to SIMR foci, dependent on HRDE-2, but only in its small RNA unbound state. This germ granule localization is critical to promote the small RNA binding specificity of HRDE-1 and, in the absence of HRDE-2, HRDE-1 exclusively loads CSR-class 22G-RNAs rather than WAGO-class 22G-RNAs, resulting in H3K9me3 deposition on CSR-target genes. Thus, our study demonstrates that HRDE-2 is critical to ensure that the correct small RNAs are used to guide nuclear RNA silencing in the C. elegans germline.

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TP12 adult with emb 3

Project description:Timepoint 12 wild type adult with emb 3 vs mixed stage reference Keywords: other

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TP11 adult with emb 2

Project description:Timepoint 11 wild type adult with emb 2 vs mixed stage reference Keywords: other

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RNA-seq of E17.5 WT and Emb-/- mouse placentas and lungs

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2023-08-31 | E-MTAB-10641 | biostudies-arrayexpress

Germ granule association drives small RNA specificity for a nuclear Argonaute protein [sRNA-seq]

Project description:RNA interference (RNAi) is a conserved gene silencing process that exists in diverse organisms to protect genome integrity and regulate gene expression. In C. elegans, majority of RNAi components are located in phase-separated perinuclear germ granules, including P granule, Mutator foci, Z granule, and SIMR foci. However, the protein components and function of the newly discovered SIMR foci are unknown. Here we identified HRDE-2 as a SIMR foci component, which interacts with the germline nuclear RNAi Argonaute HRDE-1. We found that unloaded HRDE-1 localizes to SIMR foci and this localization depends on HRDE-2. In addition, HRDE-2 contributes to HRDE-1 small RNAs binding specificity and, in the absence of HRDE-2, HRDE-1 exclusively loads CSR-class 22G-RNAs rather than WAGO-class 22G-RNAs, promotes H3K9me3 deposition on CSR-targets, but does not silence these CSR-target genes. Thus, our study demonstrates that HRDE-2 is critical to ensure correct small RNAs are used to guide nuclear RNA silencing in the C. elegans germline.

2023-08-01 | GSE239289 | GEO

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