PDF(Pubmed)
Abstract：
N-acetyltransferase 10 (NAT10)-mediated N4-acetylcytidine (ac4C) modification is crucial for mRNA stability and translation efficiency, yet the underlying function in mammalian preimplantation embryos remains unclear. Here, we characterized the ac4C modification landscape in mouse early embryos and found that the majority of embryos deficient in ac4C writer-NAT10 failed to develop into normal blastocysts. Through single-cell sequencing, RNA-seq, acetylated RNA immunoprecipitation combined with PCR (acRIP-PCR), and embryonic phenotype monitoring, Nop2 was screened as a target gene of Nat10. Mechanistically, Nat10 knockdown decreases the ac4C modification on Nop2 mRNA and reduces RNA and protein abundance by affecting the mRNA stability of Nop2. Then, depletion of NOP2 may inhibit the translation of transcription factor TEAD4, resulting in defective expression of the downstream lineage-specific gene Cdx2, and ultimately preventing blastomeres from undergoing the trophectoderm (TE) fate. However, exogenous Nop2 mRNA partially reverses this abnormal development. In conclusion, our findings demonstrate that defective ac4C modification of Nop2 mRNA hinders the morula-to-blastocyst transition by influencing the first cell fate decision in mice.
摘要：
N-乙酰转移酶10（NAT10）介导的N4-乙酰胞苷（ac4C）修饰对于mRNA的稳定性和翻译效率至关重要。然而哺乳动物胚胎植入前胚胎的潜在功能仍不清楚.这里,我们对小鼠早期胚胎中的ac4C修饰景观进行了表征，发现大多数缺乏ac4Cwriter-NAT10的胚胎未能发育成正常的胚泡。通过单细胞测序，RNA-seq,乙酰化RNA免疫沉淀结合PCR（acRIP-PCR），和胚胎表型监测，筛选Nop2作为Nat10的靶基因。机械上,Nat10敲低降低了对Nop2mRNA的ac4C修饰，并通过影响Nop2的mRNA稳定性来降低RNA和蛋白质丰度。然后，NOP2的消耗可能会抑制转录因子TEAD4的翻译，导致下游谱系特异性基因Cdx2的表达缺陷，并最终阻止卵裂球经历滋养外胚层（TE）命运。然而,外源性Nop2mRNA部分逆转了这种异常发育。总之,我们的发现表明，Nop2mRNA的ac4C修饰缺陷通过影响小鼠的第一个细胞命运决定，阻碍了桑ula到胚泡的转变。
