The role of ALKBH5 in maternal RNA decay during mammalian oocyte meiosis
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ABSTRACT:
PROVIDER: PRJNA955804 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR24175886_1.fastq.gz | Fastqsanger.gz | |||
| SRR24175886_2.fastq.gz | Fastqsanger.gz | |||
| SRR24175887_1.fastq.gz | Fastqsanger.gz | |||
| SRR24175887_2.fastq.gz | Fastqsanger.gz | |||
| SRR24175888_1.fastq.gz | Fastqsanger.gz |
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Project description:To investigate the function of ALKBH5 in the regulation of maternal RNA decay during mice oocyte meiosis, we established Alkbh5 knockout mice and analyzed the m6A level changes between control and Alkbh5 knockout oocyte at GV stage
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Project description:To investigate the function of ALKBH5 in the regulation of maternal RNA decay during mice oocyte meiosis, we established Alkbh5 knockout mice and analyzed the transcriptome changes between control and Alkbh5 knockout oocyte at different meiotic maturation stages
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Project description:The role of ALKBH5 in maternal RNA decay during mammalian oocyte meiosis [RNA-seq]
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Project description:N6-methyladenosine (m6A) is the most prevalent internal modification of messenger RNA (mRNA) in higher eukaryotes. Here we report ALKBH5 as a new mammalian demethylase that oxidatively removes the m6A modification in mRNA in vitro and inside cells. This demethylation activity of ALKBH5 significantly affects mRNA export and RNA metabolism as well as the assembly of mRNA processing factors in nuclear speckles. Alkbh5-deficient male mice are characterized by impaired fertility resulting from apoptosis that affects meiotic metaphase-stage spermatocytes. In accordance with this defect, we have identified in mouse testes 1552 differentially expressed genes which cover broad functional categories and include spermatogenesis-related mRNAs involved in the p53 functional interaction network. We show that Alkbh5-deficiency impacts the expression levels of some of these mRNAs, supporting the observed phenotype. The discovery of this new RNA demethylase strongly suggests that the reversible m6A modification plays fundamental and broad functions in mammalian cells. RNA-seq in two cell types
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Project description:In the process of mammalian maternal-to-zygotic transition (MZT), the degradation of zygotic decay (Z-decay) mRNA is later than that of maternal factor-mediated mRNA decay (M-decay) mRNA. The regulatory mechanism and physiological significance of Z-decay mRNA to maintain stability during meiosis are urgent scientific questions. In this study, we identified that PAPα worked together with poly(A)-bound RNA binding protein PABPN1 to promote the polyadenylation and translation of maternal transcripts, leading to rupture of germinal vesicles in mammalian oocytes. The oocytes specifically knocked out Pabpn1 at the primary follicle stage could develop into the fully grown (FGO) stage, but hardly could enter to the meiotic process. Furthermore, PABPN1 maintains the stability of Z-decay maternal transcripts during oocyte meiotic maturation. A number of Z-decay transcripts and the translational activity in Pabpn1-null oocytes was significantly lower than that of wild-type oocytes during meiosis. Therefore, during meiotic maturation of oocytes, PABPN1 maintains the stability of Z-decay transcripts and promotes the degradation of some M-decay transcripts, ensuring normal GVBD and polar body-1 emission.
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Project description:Oocyte meiosis is an important factor affecting female reproduction. Breast cancer amplified sequence 2 (BCAS2) is a component of the spliceosome. Previous reports have shown that BCAS2 is critical in male germ cell meiosis, oocyte development, and early embryo genome integrity. However, the role of BCAS2 in oocyte meiosis has not been reported. We used Stra8-GFP-Cre mice to knock out BCAS2 during the pachytene phase of oocytes. The results of fertility tests showed that the cko mice were infertile. Morphological analysis showed that the number of primary follicles of 2M ovary was significantly reduced and follicle development was blocked. Further analysis showed that the number of primordial follicles decreased and follicle development slowed from 7dpp ovaries. Sequencing revealed that DNA damage in oocytes could not be repaired from 5dpp. There was an abnormality in meiosis, some oocytes could not reach the diplotene stage of meiosis, and more oocytes could not develop to the dictyate stage. AS analysis reveals that abnormal variable splicing of Dazl and Diaph2 Oogens-related genes in cKO mice, with involvement of the PRP19/CDC5l complex.
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Project description:The faithful execution of molecular program underlying oocyte maturation and meiosis is vital to generate competent haploid gametes for effective reproduction in mammals. However, the organization and principle of gene modules and molecular circuits for oocyte meiosis remain obscure. Here, we employed the recently developed single cell RNA-seq technique to profile the transcriptomes of germinal vesicle (GV) and metaphase II (MII) oocytes, aiming to discover the dynamic changes of mRNAs and long non-coding RNAs (lncRNAs) during oocyte meiotic maturation. During the transition from GV to MII, total number of RNAs (mRNAs and lncRNAs) in oocytes decreased. Moreover, 1807 (602 up- and 1205 down-regulated) mRNAs and 313 (177 up- and 136 down-regulated) lncRNAs were significantly differentially expressed. During meiotic maturation, expression level of most mRNAs decreased, but mitochondrial RNAs, and also more lncRNAs and their target genes increased. Both DE mRNAs and lncRNAs were enriched in multiple biological and signal pathways potentially associated with oocyte meiosis. Highly abundantly expressed mRNAs (including DNMT1, UHRF2, PCNA, ARMC1, BTG4, ASNS and SEP11) and lncRNAs were also discovered. Weighted gene co-expression network analysis (WGCNA) revealed 20 hub mRNAs, and three modules of contrasting functions in oocyte maturation and meiosis. Taken together, our findings provide insights and resources for further functional investigation of mRNAs/lncRNAs in mammalian oocyte meiosis and maturation.
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Project description:During Caenorhabditis elegans oocyte meiosis, a multi-protein complex localised between homologous chromosomes, the ring complex (RC), promotes chromosome congression through the action of the chromokinesin KLP-19. While some RC components are known, the mechanism of RC assembly has remained obscure. A germline-specific screen identified KLP-19 as a SUMO substrate in vivo and we found that the SUMO E3 ligase GEI-17/PIAS is required for KLP-19 recruitment to the RC. Additionally, KLP-19 is efficiently sumoylated in vitro in a GEI-17-dependent manner. Further biochemical analysis showed that KLP-19 and GEI-17 are efficiently modified by SUMO and that GEI-17 and another RC component, the kinase BUB-1, can interact non-covalently with SUMO. While SUMO conjugation is required for RC assembly, we also provide evidence consistent with non-covalent SUMO interactions contributing to RC assembly in vivo. Our results highlight the importance of sumoylation and non-covalent SUMO interaction in regulating dynamic protein complex assembly during oocyte meiosis.
2020-10-15 | PXD005202 | Pride
Project description:A decrease in oocyte developmental potential is a major obstacle for successful pregnancy in women of advanced age. However, the age-related epigenetic modifications associated with dynamic transcriptome changes, particularly meiotic maturation-coupled mRNA clearance, have not been adequately characterized in human oocytes. This study demonstrate a decreased storage of transcripts encoding key factors regulating the maternal mRNA degradome in fully grown oocytes of women of advanced age. A similar defect in meiotic maturation-triggered mRNA clearance was also detected in aged mouse oocytes. Mechanistically, the epigenetic and cytoplasmic aspects of oocyte maturation are synchronized in both the normal development and aging processes. The level of histone H3K4 trimethylation (H3K4me3) was high in fully grown mouse and human oocytes derived from young females but decreased during aging due to the decreased expression of epigenetic factors responsible for H3K4me3 accumulation. Oocyte-specific knockout of the gene encoding CxxC-finger protein 1 (CXXC1), a DNA-binding subunit of SETD1 methyltransferase, caused ooplasm changes associated with accelerated aging and impaired maternal mRNA translation and degradation. These results suggest that a network of CXXC1-maintained H3K4me3, in association with mRNA decay competence, sets a timer for oocyte deterioration and plays a role in oocyte aging in both mouse and human oocytes.
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