Mus musculus strain:C57BL/6
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA634688 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR11881000.fastq.gz | Fastqsanger.gz | |||
| SRR11881001.fastq.gz | Fastqsanger.gz | |||
| SRR11881002.fastq.gz | Fastqsanger.gz | |||
| SRR11881003.fastq.gz | Fastqsanger.gz | |||
| SRR11881004.fastq.gz | Fastqsanger.gz |
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Project description:PIWI-interacting RNAs(piRNAs) are required for transposon repression, de novo DNA methylation, epigenetic and post-transcriptional regulation of gene expression in the germline. A mist of piRNA biogenesis is the identification of the 3’-5’ exnuclease that trimms the 3’ end of piRNA intermediates to form mature piRNAs in mammals. Here, we show that mice deficient for PNLDC1, an evolutionarily conserved 3’-5’ exonuclease, accumulate 3’ untrimmed piRNA intermediates of 30~40nt. These 3’ extended piRNA intermediates associate with PIWI proteins, but are impaired for function. Male Pnldc1 knockout mice display LINE1 derepression and are sterile owing to chimeric spermatogenetic arrest at spermatocyte and spermatid stages. Our findings illustrate the conserved role of mouse Pnldc1 in 3’ trimming during piRNA maturation. However, since Pnldc1 knockout in mice creates phenotypes distinct from its reported interactive partner, Tdrkh, the panorama of participants in mammalian piRNA 3’ end maturation may be more complex than previously speculated.
2021-02-23 | E-MTAB-8335 | biostudies-arrayexpress
Project description:PIWI-interacting RNAs(piRNAs) are required for transposon repression, de novo DNA methylation, epigenetic and post-transcriptional regulation of gene expression in the germline. A mist of piRNA biogenesis is the identification of the 3’-5’ exnuclease that trimms the 3’ end of piRNA intermediates to form mature piRNAs in mammals.Here, we show that mice deficient for PNLDC1, an evolutionarily conserved 3’-5’ exonuclease, accumulate 3’ untrimmed piRNA intermediates of 30~40nt. These 3’ extended piRNA intermediates associate with PIWI proteins, but are impaired for function. Male Pnldc1 knockout mice display LINE1 derepression and are sterile owing to chimeric spermatogenetic arrest at spermatocyte and spermatid stages. Our findings illustrate the conserved role of mouse Pnldc1 in 3’ trimming during piRNA maturation. However, since Pnldc1 knockout in mice creates phenotypes distinct from its reported interactive partner,Tdrkh,the panorama of participants in mammalian piRNA 3’ end maturation may be more complex than previously speculated .There is the sequence of Pnldc1 Total RNA-seq of P10 Testis.
2022-08-17 | E-MTAB-9486 | biostudies-arrayexpress
Project description:The piRNA pathway is an adaptive mechanism for maintaining the stability of the genome by repression selfish genomic elements such as transposons. As a host defense mechanism the piRNA pathway is rapidly evolving with different sets of piRNA silencing different targets in closely-related species. In male germline of D.melanogaster, repression of Stellate genes by piRNA generated from Supressor of Stellate, Su(Ste), locus is required for male fertility, but both Su(Ste) piRNA and their targets are absent in other Drosophila species. Here we showed that D.melanogaster genome contain non-coding genomic repeats that have sequence similarity to vasa, protein-coding host gene essential for germline development. In male germline Vasa Related (Varel) repeats produce abundant piRNA that are anti-sense to vasa, however, vasa mRNA escape silencing due to imperfect complementarity between piRNA and vasa sequences. Unexpectedly, we found that Varel piRNA of D.melanogaster target vasa of closely related D. mauritana species in interspecies hybrids. The same hybrids show derepression of harmful Stellate locus due to the absence of Su(Ste) piRNA. Thus, the piRNA pathway contributes to the reproductive isolation between Drosophila species, causing hybrid male sterility via misregulation of two different host factors.
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Project description:In the male mouse germline, PIWI-interacting RNAs (piRNAs), bound by the PIWI protein MIWI2 (PIWIL4), guide DNA methylation of young active transposons through SPOCD1. However, the underlying mechanisms of SPOCD1-mediated piRNA-directed transposon methylation and whether this pathway functions to protect the human germline remains unknown. We identified loss-of-function variants in human SPOCD1 that cause defective transposon silencing and male infertility. Through the analysis of one of these pathogenic alleles, we discovered that the uncharacterised protein C19ORF84 interacts with SPOCD1. DNMT3C, the DNA methyltransferase responsible for transposon methylation, associates with SPOCD1 and C19ORF84 in foetal gonocytes. Furthermore, C19ORF84 is essential for piRNA-directed DNA methylation and male mouse fertility. Finally, C19ORF84 mediates the in vivo association of SPOCD1 with the de novo methylation machinery. In summary, we have discovered a conserved role for the human piRNA pathway in transposon silencing and C19ORF84, an uncharacterised protein essential for orchestrating piRNA-directed DNA methylation.
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Project description:In diverse organisms, the highly-conserved Piwi proteins bind to a complex class of small non-coding RNAs called piRNAs, and are essential for germline stem cell maintenance, transposon silencing, fertility, and offspring viability1. By homology with other Argonautes, Piwi proteins have been proposed and later reported to possess endonuclease activity in vitro, effected by its conserved aspartate catalytic triad2,3. Indeed, mutation of these residues in one of the three mouse PIWI proteins, MILI, affects the production of Line 1 piRNAs and spermatogenesis4. In Drosophila, the conserved slicer function has been proposed to be crucial for all Piwi proteins in piRNA amplification and transposon silencing5. Here, we report in vivo evidence that, in contrast to common belief, the catalytic triad of Piwi is not required for the function of PIwi in the germline for stem cell maintenance, fertility, transposon silencing, or piRNA biogenesis. In addition, it is not required for piRNA biogenesis in the soma where Piwi is the sole Piwi-subfamily protein involved in the process. These observations, together with the recent findings on MILI and MIWI2 triad mutant, indicate that even though the slicer function may be required for a subset of piRNA-mediated process, it is overall inconsequential for the function of Piwi proteins Assess the role of the endonuclease activity of Piwi in all known biological processes that involve Piwi function in Drosophila ovary
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Project description:In the fetal mouse testis, PIWI Interacting RNAs (piRNAs) guide PIWI proteins to silence transposons, but after birth, most post-pubertal pachytene piRNAs map to genome uniquely and are thought to regulate genes required for male fertility. In human males, the developmental classes, precise genomic origins, and transcriptional regulation of post-natal piRNAs remain undefined. Here, we demarcate the genes and transcripts that produce post-natal piRNAs in human juvenile and adult testes. As in mouse, A‑MYB in humans drives transcription of both pachytene piRNA precursor transcripts and the mRNAs encoding piRNA biogenesis factors. Although human piRNA genes are syntenic to those in other placental mammals, their sequences are poorly conserved. In fact, pachytene piRNA loci are rapidly diverging even among modern humans. Our findings suggest that during mammalian evolution, pachytene piRNA genes are under fewer selective constraints. We speculate that pachytene piRNA diversity may provide a hitherto unrecognized driver of reproductive isolation.
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Project description:Purpose: piRNAs are a class of germline-specific, small non-coding RNAs required to maintain genome integrity and preserve RNA homeostasis essential for male gametogenesis. During mouse spermatogenesis, male germ cells transcribe piRNAs at pre-pachytene anad pachytene stages of development from genetically distince loci. The most well-studied and conserved function of pre-pachytene piRNAs is repression of transposons to ensure the integrity of the germline genome. In contrast to pre-pachytene piRNAs, the mode of action and function of pachytene piRNA is much less understood. To investigate and characterize the functional importance of pachytene piRNAs in spermatogenesis and male fertility, we have used CRISPR/Cas9 and established mutant mice which have deleted a bi-directional promoter of the pachytene piRNA cluster on Chr18. Mutant mice have a male sterile phenotype with acrosome overgrowth and defective motility.
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Project description:Piwi proteins and Piwi-interacting RNAs (piRNAs) are best known for their roles in suppressing transposons and promoting fertility. Yet piRNA biogenesis and its mechanisms of action differ widely between distantly related species. To better understand the evolution of piRNAs, we characterized the piRNA pathway in C. briggsae, a sibling species of the model organism C. elegans. Our analyses define 25,883 piRNA producing-loci in C. briggsae. piRNA sequences in C. briggsae are extremely divergent from their counterparts in C. elegans, yet both species adopt similar genomic organization and transcription program that drive piRNA expression. By examining production of Piwi-dependent secondary small RNAs, we identified a set of protein-coding genes that are evolutionarily conserved piRNA targets. In contrast to C. elegans, small RNAs mapped to ribosomal RNAs or histone transcripts are not hyper-accumulated in C. briggsae. Instead, we found that fewer introns in transcripts are associated with hyper-accumulation of small RNAs. Together our work highlights evolutionary conservation and divergence of the nematode piRNA pathway and provides insights into its role in endogenous gene regulation.
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