Bombyx mori histone methyltransferase BmAsh2 is essential for silkworm piRNA-mediated sex determination.

Bombyx mori histone methyltransferase BmAsh2 is essential for silkworm piRNA-mediated sex determination.
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家蚕组蛋白甲基转移酶 BmAsh2 对于家蚕 piRNA 介导的性别决定至关重要。

DOI:
10.1371/journal.pgen.1007245
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发表时间:
2018-03
期刊:
影响因子:
4.5
通讯作者:
Tan A
Tan A
中科院分区:
生物学2区
文献类型:
--
作者:
Li Z;You L;Yan D;James AA;Huang Y;Tan A

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性别决定是一个高度多样性的分级调控过程,在包括昆虫在内的不同生物中存在。W染色体来源的Fem皮尔纳已被鉴定为鳞翅目昆虫桑尺蠖(Escherichia mori)中的主要性别决定因子,揭示了独特的piRNA介导的性别决定途径。然而,家蚕性别决定的综合机制仍然知之甚少。我们在这里表明,家蚕PIWI蛋白BmSiwi,而不是BmAgo 3,是必不可少的家蚕性别决定。CRISPR/Cas9介导的BmSiwi耗竭导致卵子发生中的发育停滞和部分雌性性逆转,而BmAgo 3耗竭仅影响卵子发生。我们确定了三个组蛋白甲基转移酶(HMTs),显着下调BmSiwi突变蛾。其中之一BmAsh 2的破坏会导致piRNA和转座因子(TE)的失调,支持它在皮尔纳信号通路中的作用。更重要的是,我们发现BmAsh 2诱变导致卵子发生停滞和部分女性到男性的性逆转以及性别决定基因Bmdsx和BmMasc的失调。其他两种HMT BmSETD 2和BmEggless的突变不影响piRNA介导的性别决定。组织学分析和免疫沉淀结果支持BmAsh 2和BmSiwi蛋白之间的功能性相互作用。我们的数据提供了HMT BmAsh 2在家蚕piRNA介导的性别决定中起关键作用的第一个证据。
Sex determination is a hierarchically-regulated process with high diversity in different organisms including insects. The W chromosome-derived Fem piRNA has been identified as the primary sex determination factor in the lepidopteran insect, Bombyx mori, revealing a distinctive piRNA-mediated sex determination pathway. However, the comprehensive mechanism of silkworm sex determination is still poorly understood. We show here that the silkworm PIWI protein BmSiwi, but not BmAgo3, is essential for silkworm sex determination. CRISPR/Cas9-mediated depletion of BmSiwi results in developmental arrest in oogenesis and partial female sexual reversal, while BmAgo3 depletion only affects oogenesis. We identify three histone methyltransferases (HMTs) that are significantly down-regulated in BmSiwi mutant moths. Disruption one of these, BmAsh2, causes dysregulation of piRNAs and transposable elements (TEs), supporting a role for it in the piRNA signaling pathway. More importantly, we find that BmAsh2 mutagenesis results in oogenesis arrest and partial female-to-male sexual reversal as well as dysregulation of the sex determination genes, Bmdsx and BmMasc. Mutagenesis of other two HMTs, BmSETD2 and BmEggless, does not affect piRNA-mediated sex determination. Histological analysis and immunoprecipitation results support a functional interaction between the BmAsh2 and BmSiwi proteins. Our data provide the first evidence that the HMT, BmAsh2, plays key roles in silkworm piRNA-mediated sex determination.
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