PABPN1 promotes meiotic maturation of oocytes by interacting with PAPOLA
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA762877 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR15871701_1.fastq.gz | Fastqsanger.gz | |||
| SRR15871701_2.fastq.gz | Fastqsanger.gz | |||
| SRR15871702_1.fastq.gz | Fastqsanger.gz | |||
| SRR15871702_2.fastq.gz | Fastqsanger.gz | |||
| SRR15871703_1.fastq.gz | Fastqsanger.gz |
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Project description:Poly(A) polymerase α (PAPα), as the specific mRNA polyadenylation enzyme in the cytoplasm of mammalian oocytes, is essential for oocytes to exclude the first polar body. However, PAPα knockout did not affect germinal vesicles breakdown (GVBD) of oocytes, and the mechanism needs to be further explored. In this study, we identified that PAPα work together with poly(A)-bound RNA binding protein PABPN1 to promote the rupture of germinal vesicles in mammalian oocytes. The protein level of Pabpn1 gradually increases with the meiotic maturation of oocytes. The oocytes specifically knocked out Pabpn1 at the primary follicle stage could develop into the fully grown (FGO) stage, but hardly could enter into the meiotic process. The activated form of CDK1 was injected into Pabpn1-null oocytes, the oocytes could enter into meiotic process. The translational activity of Pabpn1-null oocytes was significantly lower than that of wild-type oocytes during meiosis. In particular, the expression level of protein (BTG4 and CDC25), which were essential for the meiotic maturation of oocytes, were significantly decreased. Therefore, during the oocyte meiosis process, PABPN1 and PAPα jointly promote the oocyte to enter the meiosis process.
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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:The growing oocytes stored a large amount of maternal mRNA to support the subsequent "maternal-zygotic transition" process, which is a key regulatory events in female reproduction. The accumulation of maternal mRNA in the growing oocyte determines the developmental potential of the oocyte. At present, it is not clear how the growing oocytes store and process the newly transcribed mRNA under physiological conditions, including the distribution and localization of mRNA in the oocyte, as well as the molecular mechanism and physiological significance of this localization. In this study, we found that poly(A) tailed mRNA has the dynamic distributions of droplet fusion and gradual disappearance in the mouse nucleus as oocyte growth and meiotic maturation. Our research found that PABPN1 modulated the dynamic compartmentalization of mRNA with poly(A) tails to form NmPD (nuclear mRNA processing domain) during the growing oocytes through phase separation. The NmPD in Pabpn1-null oocytes could not form the compartmentalized distribution, and the Pabpn1 knockout females were sterile with defects of premature ovarian failure. Combined with multi-omics sequencing analysis, we investigate that NmPD was essential for long 3'-UTR isoform formation and the stability of mRNA in growing oocytes.
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Project description:Identification of PABPN1 interacting proteins by proximity labeling with TurboID coupled with mass spectrometry.
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Project description:Spermidine is involved in a variety of biological processes, including DNA metabolism, autophagy and aging. Previous studies have shown that spermidine can increase the percentage of mouse oocytes developing into blastocysts after in vitro fertilization. However, none of the past studies elucidated the effects of spermidine supplementation on porcine oocyte maturation. Here, we choose appropriate dose of spermidine to be added to the maturation medium during in vitro maturation (IVM) to verify whether spermidine can actively promote the maturation of porcine oocytes. Our study provided substantial evidence that spermidine exposure promoted the porcine oocyte meiotic maturation. In addition, single-cell transcriptome analysis identified the target effectors of spermidine actions in porcine oocytes, further demonstrating that spermidine exposure enhanced mitochondrial distribution and function, leading to a reduced excessive oxidative stress-induced DNA damage and early apoptosis of porcine oocytes. These findings demonstrate that spermidine not only delays ageing and cancer treatment, but also improves the quality of germ cells, which may be less likely to suffer from sterility or infertility in humans and animals.
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Project description:The polyadenylation of mRNA is pivotal for mRNA stability and efficient mRNA translation. The length of poly(A) tail is dynamically changed under various physiological conditions; however, the functions and mechanisms of aberrant poly(A) length control in cancers remain poorly understood. Here we uncovered the aberrant lengthening of poly(A) tail and particularly upregulated expression of poly(A) polymerase alpha (PAPOLA) in acute myeloid leukemia (AML), and the elevated PAPOLA expression significantly correlated with unfavorable AML outcomes. We further demonstrated the critical oncogenic functions of PAPOLA-mediated poly(A) lengthening in promoting AML leukemogenesis and leukemia stem cell self-renewal using human primary AML samples, AML cells and various mouse leukemogenesis models. Mechanistically, we identified GSTM2 as a key downstream target of PAPOLA that regulated metabolic reprogramming through the HNE-DLD axis to promote AML initiation and progression. Moreover, PAPOLA inhibitor cordycepin effectively blocked metabolic reprogramming and AML leukemogenesis in vivo. Overall, our study uncovers the novel functional link between aberrant poly(A) lengthening and cellular metabolic reprogramming in AML, and provides a molecular basis for development of an effective therapeutic strategy for AML patients.
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Project description:Mammalian oocytes establish a unique landscape of histone modifications, some of which are inherited by early embryos. How histone variants shape the maternal histone landscape remains unknown. Here, we map histone H2A variants in mouse fully grown oocytes (FGOs) and find that H2A.Z forms broad domains across intergenic regions, along non-canonical H3K4me3 (ncH3K4me3). During oocyte growth, H2A.Z progressively transitions from an active promoter-rich, canonical distribution to a non-canonical broad distribution (ncH2A.Z). Depletion of H2A.Z in oocytes partially impairs ncH3K4me3 formation and causes severe defects in meiotic progression, which resemble Mll2-knockout oocytes. Conversely, depletion of ncH3K4me3 by Mll2 knockout also causes a reduction of ncH2A.Z in FGOs. Thus, our study suggests that ncH2A.Z and ncH3K4me3 reinforce each other to form functional oocytes.
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