ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/013/SRR27737113/SRR27737113_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/009/SRR27737109/SRR27737109_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/010/SRR27737110/SRR27737110_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/013/SRR27737113/SRR27737113_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/014/SRR27737114/SRR27737114_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/011/SRR27737111/SRR27737111_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/009/SRR27737109/SRR27737109_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/014/SRR27737114/SRR27737114_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/012/SRR27737112/SRR27737112_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/010/SRR27737110/SRR27737110_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/012/SRR27737112/SRR27737112_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR277/011/SRR27737111/SRR27737111_1.fastq.gzprimaryOK200GenomicsFan heng-yu, Life Sciences Institutehttps://www.ebi.ac.uk/ena/browser/view/PRJNA1069628Mus musculusxref:PubMed:39982985Mammalian oocyte maturation is driven by strictly translational regulation of maternal mRNAs stored in the cytoplasm. However, the function and mechanism of post-transcriptional chemical modifications especially the newly identified N4-acetylcytidine (ac4C) catalyzed by N-acetyltransferase 10 (NAT10) in this process are previously unknown. In this study, we developed a low-input ac4C sequencing technology—ac4C LACE-seq and mapped 8241 ac4C peaks at the whole transcriptome level using 50 mouse oocytes at the germinal vesicle (GV) stage. We profiled the mRNA landscapes of NAT10-interactions and ac4C modifications. The NAT10-interacted and ac4C modified transcripts displayed association with high translation efficiency in oocytes. Oocyte-specific Nat10 knockout wiped out ac4C signals in oocytes and caused severe defects in meiotic maturation and female infertility. ac4C LACE-seq results indicated that Nat10 deletion led to a failure of ac4C deposition on mRNAs encoding key maternal factors such as MAY2, ZAR1, BTG4 and cyclin B1 that regulate transcriptome stability and maternal-to-zygotic transition. Nat10-deleted oocytes had decreased mRNA translation efficiencies during meiotic maturation, partially due to the direct inhibition ac4C sites on specific transcripts. In sum, we developed low-input, high-sensitivity mRNA ac4C profiling approach and highlighted the important physiological function of ac4C in precise regulation of the oocyte meiotic maturation by enhancing translation efficiency. Overall design: The oocytes were harvested from mature WT and Gdf9-cre Nat10 conditional knockout female mice 32-48h after PMSG and washed by 0.2% containing PBS for 3 times, then 10 oocytes per sample were lysed in lysis buffer (containing 0.2% Triton X-100 and Recombination RNase Inhibitor) and prceeded for cDNA library constribution following the workflow of Smart-seq2. And for each sample, ERCC (1:1000, 0.2μl/sample) were added as spike-in for further calibration.ENAfalseNAT10-mediated mRNA N4-acetylation is Essential for the Translational Regulation During Oocyte Meiotic Maturation in MiceNAT10-mediated mRNA N4-acetylation is Essential for the Translational Regulation During Oocyte Meiotic Maturation in Mice2025-09-242024-02-20PRJNA1069628GSE2542881009039982985