Effects of RNAi-mediated knockdown of histone methyltransferases on the sex-specific mRNA expression of Imp in the silkworm Bombyx mori.
Effects of RNAi-mediated knockdown of histone methyltransferases on the sex-specific mRNA expression of Imp in the silkworm Bombyx mori.
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DOI：
10.3390/ijms15046772
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发表时间：
2014-04-22
影响因子：
5.6
通讯作者：
Aoki F
中科院分区：
文献类型：
--
作者：
Suzuki MG;Ito H;Aoki F
家蚕的性别分化是由 Bmdsx 的性别特异性剪接控制的，这导致外显子 3 和 4 以雄性特异性方式缺失。在家蚕中，胰岛素样生长因子 II mRNA 结合蛋白 (Imp) 是参与 Bmdsx 雄性特异性剪接的雄性特异性因子。男性特异性Imp mRNA源自男性特异性外显子8的包含。为了验证Imp中组蛋白甲基化和替代RNA加工之间的联系，我们检查了RNAi介导的几种组蛋白甲基转移酶的敲低对Imp的性别特异性mRNA表达的影响。结果，当 H3K79 甲基转移酶 DOT1L 的表达被抑制到 < 对照男性的 10% 时，Imp mRNA 的男性特异性表达被完全消除。染色质免疫沉淀定量 PCR 分析显示，Imp 中 H3K79me2 在正常男性中的分布高于正常女性。 RNA 聚合酶 II (RNAP II) 持续合成能力测定表明，与对照男性相比，雄性 DOT1L 的 RNAi 敲低导致 RNAP II 持续合成能力下降两倍，几乎与正常女性中观察到的水平相同。抑制雄性细胞中 RNAP II 介导的伸长会抑制 Imp 的雄性特异性剪接。我们的数据表明，H3K79me2 沿着 Imp 的积累可能与由于 RNAP II 持续合成能力增加而导致的 Imp mRNA 的男性特异性替代加工有关。
Sexual differentiation in Bombyx mori is controlled by sex-specific splicing of Bmdsx, which results in the omission of exons 3 and 4 in a male-specific manner. In B. mori, insulin-like growth factor II mRNA-binding protein (Imp) is a male-specific factor involved in male-specific splicing of Bmdsx. Male-specific Imp mRNA results from the male-specific inclusion of exon 8. To verify the link between histone methylation and alternative RNA processing in Imp, we examined the effects of RNAi-mediated knockdown of several histone methyltransferases on the sex-specific mRNA expression of Imp. As a result, male-specific expression of Imp mRNA was completely abolished when expression of the H3K79 methyltransferase DOT1L was repressed to <10% of that in control males. Chromatin immunoprecipitation-quantitative PCR analysis revealed a higher distribution of H3K79me2 in normal males than in normal females across Imp. RNA polymerase II (RNAP II) processivity assays indicated that RNAi knockdown of DOT1L in males caused a twofold decrease in RNAP II processivity compared to that in control males, with almost equivalent levels to those observed in normal females. Inhibition of RNAP II-mediated elongation in male cells repressed the male-specific splicing of Imp. Our data suggest the possibility that H3K79me2 accumulation along Imp is associated with the male-specific alternative processing of Imp mRNA that results from increased RNAP II processivity.
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影响因子：
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作者：
Milcarek, Christine;Albring, Michael;Park, Kyung Soo
通讯作者：
Park, Kyung Soo
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EXINGER, F;LACROUTE, F
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LACROUTE, F
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Kolasinska-Zwierz P;Down T;Latorre I;Liu T;Liu XS;Ahringer J
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Misteli T
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作者：
de la Mata, Manuel;Lafaille, Celina;Kornblihtt, Alberto R.
通讯作者：
Kornblihtt, Alberto R.