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Multi-omics analyses demonstrate a critical role for EHMT1 methyltransferase in transcriptional repression during oogenesis
Genome Research ( IF 7 ) Pub Date : 2023-01-01 , DOI: 10.1101/gr.277046.122 Hannah Demond 1, 2 , Courtney W Hanna 1, 3, 4 , Juan Castillo-Fernandez 1 , Fátima Santos 1, 3 , Evangelia K Papachristou 5 , Anne Segonds-Pichon 6 , Kamal Kishore 5 , Simon Andrews 6 , Clive S D'Santos 5 , Gavin Kelsey 1, 3, 7
Genome Research ( IF 7 ) Pub Date : 2023-01-01 , DOI: 10.1101/gr.277046.122 Hannah Demond 1, 2 , Courtney W Hanna 1, 3, 4 , Juan Castillo-Fernandez 1 , Fátima Santos 1, 3 , Evangelia K Papachristou 5 , Anne Segonds-Pichon 6 , Kamal Kishore 5 , Simon Andrews 6 , Clive S D'Santos 5 , Gavin Kelsey 1, 3, 7
Affiliation
EHMT1 (also known as GLP) is a multifunctional protein, best known for its role as an H3K9me1 and H3K9me2 methyltransferase through its reportedly obligatory dimerization with EHMT2 (also known as G9A). Here, we investigated the role of EHMT1 in the oocyte in comparison to EHMT2 using oocyte-specific conditional knockout mouse models (Ehmt2 cKO, Ehmt1 cKO, Ehmt1/2 cDKO), with ablation from the early phase of oocyte growth. Loss of EHMT1 in Ehmt1 cKO and Ehmt1/2 cDKO oocytes recapitulated meiotic defects observed in the Ehmt2 cKO; however, there was a significant impairment in oocyte maturation and developmental competence in Ehmt1 cKO and Ehmt1/2 cDKO oocytes beyond that observed in the Ehmt2 cKO. Consequently, loss of EHMT1 in oogenesis results, upon fertilization, in mid-gestation embryonic lethality. To identify H3K9 methylation and other meaningful biological changes in each mutant to explore the molecular functions of EHMT1 and EHMT2, we performed immunofluorescence imaging, multi-omics sequencing, and mass spectrometry (MS)–based proteome analyses in cKO oocytes. Although H3K9me1 was depleted only upon loss of EHMT1, H3K9me2 was decreased, and H3K9me2-enriched domains were eliminated equally upon loss of EHMT1 or EHMT2. Furthermore, there were more significant changes in the transcriptome, DNA methylome, and proteome in Ehmt1/2 cDKO than Ehmt2 cKO oocytes, with transcriptional derepression leading to increased protein abundance and local changes in genic DNA methylation in Ehmt1/2 cDKO oocytes. Together, our findings suggest that EHMT1 contributes to local transcriptional repression in the oocyte, partially independent of EHMT2, and is critical for oogenesis and oocyte developmental competence.
中文翻译:
多组学分析表明 EHMT1 甲基转移酶在卵子发生过程中的转录抑制中起关键作用
EHMT1(也称为 GLP)是一种多功能蛋白,以其作为 H3K9me1 和 H3K9me2 甲基转移酶的作用而闻名,据报道它通过与 EHMT2(也称为 G9A)发生强制性二聚化。在这里,我们使用卵母细胞特异性条件敲除小鼠模型(Ehmt2 cKO、Ehmt1 cKO、Ehmt1/2 cDKO)研究了 EHMT1 与 EHMT2 在卵母细胞中的作用,并从卵母细胞生长的早期阶段进行了消融。Ehmt1 cKO 和Ehmt1/2 cDKO 卵母细胞中 EHMT1 的缺失重现了在Ehmt2 cKO 中观察到的减数分裂缺陷;然而,在Ehmt1 cKO 和Ehmt1/2 cDKO 卵母细胞超出了在Ehmt2 cKO 中观察到的卵母细胞。因此,在受精后,卵子发生过程中 EHMT1 的缺失会导致妊娠中期胚胎死亡。为了识别每个突变体中的 H3K9 甲基化和其他有意义的生物学变化以探索 EHMT1 和 EHMT2 的分子功能,我们在 cKO 卵母细胞中进行了免疫荧光成像、多组学测序和基于质谱 (MS) 的蛋白质组分析。尽管 H3K9me1 仅在丢失 EHMT1 时耗尽,但 H3K9me2 减少,并且在丢失 EHMT1 或 EHMT2 时同样消除了 H3K9me2 富集域。此外, Ehmt1/2 cDKO 的转录组、DNA 甲基化组和蛋白质组的变化比Ehmt2更显着cKO 卵母细胞,转录去抑制导致蛋白质丰度增加和Ehmt1/2 cDKO 卵母细胞中基因 DNA 甲基化的局部变化。总之,我们的研究结果表明,EHMT1 有助于卵母细胞的局部转录抑制,部分独立于 EHMT2,并且对卵子发生和卵母细胞发育能力至关重要。
更新日期:2023-01-01
中文翻译:
多组学分析表明 EHMT1 甲基转移酶在卵子发生过程中的转录抑制中起关键作用
EHMT1(也称为 GLP)是一种多功能蛋白,以其作为 H3K9me1 和 H3K9me2 甲基转移酶的作用而闻名,据报道它通过与 EHMT2(也称为 G9A)发生强制性二聚化。在这里,我们使用卵母细胞特异性条件敲除小鼠模型(Ehmt2 cKO、Ehmt1 cKO、Ehmt1/2 cDKO)研究了 EHMT1 与 EHMT2 在卵母细胞中的作用,并从卵母细胞生长的早期阶段进行了消融。Ehmt1 cKO 和Ehmt1/2 cDKO 卵母细胞中 EHMT1 的缺失重现了在Ehmt2 cKO 中观察到的减数分裂缺陷;然而,在Ehmt1 cKO 和Ehmt1/2 cDKO 卵母细胞超出了在Ehmt2 cKO 中观察到的卵母细胞。因此,在受精后,卵子发生过程中 EHMT1 的缺失会导致妊娠中期胚胎死亡。为了识别每个突变体中的 H3K9 甲基化和其他有意义的生物学变化以探索 EHMT1 和 EHMT2 的分子功能,我们在 cKO 卵母细胞中进行了免疫荧光成像、多组学测序和基于质谱 (MS) 的蛋白质组分析。尽管 H3K9me1 仅在丢失 EHMT1 时耗尽,但 H3K9me2 减少,并且在丢失 EHMT1 或 EHMT2 时同样消除了 H3K9me2 富集域。此外, Ehmt1/2 cDKO 的转录组、DNA 甲基化组和蛋白质组的变化比Ehmt2更显着cKO 卵母细胞,转录去抑制导致蛋白质丰度增加和Ehmt1/2 cDKO 卵母细胞中基因 DNA 甲基化的局部变化。总之,我们的研究结果表明,EHMT1 有助于卵母细胞的局部转录抑制,部分独立于 EHMT2,并且对卵子发生和卵母细胞发育能力至关重要。