20E-mediated regulation of BmKr-h1 by BmKRP promotes oocyte maturation.
20E-mediated regulation of BmKr-h1 by BmKRP promotes oocyte maturation.
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BmKRP 20E 介导的 BmKr-h1 调节促进卵母细胞成熟
DOI：
10.1186/s12915-021-00952-2
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发表时间：
2021-02-25
期刊：
影响因子：
5.4
通讯作者：
Deng H
中科院分区：
文献类型：
--
作者：
Zhu Z;Tong C;Qiu B;Yang H;Xu J;Zheng S;Song Q;Feng Q;Deng H
Krüppel同源物1（Kr-h1）是保幼激素（JH）信号传导的关键转录因子，已知在调节昆虫变态和成虫生殖方面发挥关键作用。蜕皮激素20-羟基蜕皮激素（20-hydroxyecdysone，20 E）也能诱导Kr-h1的表达，但20 E诱导Kr-h1表达的机制尚不清楚。本研究探讨了20 E对鳞翅目模式昆虫家蚕生殖系统Kr-h1基因表达的诱导作用及其分子机制。发育和组织特异性表达分析表明，BmKr-h1在蛹后期和成虫期卵巢中高表达，且表达受20 E诱导。RNA干扰（RNAi）介导的BmKr-h1缺失严重抑制了卵母细胞中卵黄原蛋白受体（VgR）的转录，导致卵母细胞中卵黄原蛋白（Vg）沉积减少。BmKr-h1特异性结合BmVgR上游-2818至-2805 nt之间的Kr-h1结合位点（KBS），并增强BmVgR的转录。在BmKr-h1基因启动子区发现了一个20 E顺式调控元件（cis-regulatory element，CRE），并通过荧光素酶报告基因分析、EMSA和DNA-ChIP进行了功能验证。利用pull-down分析，我们鉴定了一种新的转录因子B。mori Kr-h1调节蛋白（BmKRP），其特异性结合BmKr-h1 CRE并激活其转录。CRISPR/Cas9介导的BmKRP敲除抑制了BmKr-h1和BmVgR的转录，导致卵子发生停滞。我们发现BmKRP是一个新的转录因子，介导20 E对B的调控。桑卵发生我们的数据表明，20 E诱导BmKRP调节BmKr-h1的表达，这反过来又诱导BmVgR的表达，以促进Vg摄取和卵子发生。在线版本包含补充材料，可通过10.1186/s12915-021-00952-2获得。
Krüppel homolog 1 (Kr-h1) is a critical transcription factor for juvenile hormone (JH) signaling, known to play a key role in regulating metamorphosis and adult reproduction in insects. Kr-h1 can also be induced by molting hormone 20-hydroxyecdysone (20E), however, the underlying mechanism of 20E-induced Kr-h1 expression remains unclear. In the present study, we investigated the molecular mechanism of Kr-h1 induction by 20E in the reproductive system of a model lepidopteran insect, Bombyx mori. Developmental and tissue-specific expression analysis revealed that BmKr-h1 was highly expressed in ovaries during the late pupal and adult stages and the expression was induced by 20E. RNA interference (RNAi)-mediated depletion of BmKr-h1 in female pupae severely repressed the transcription of vitellogenin receptor (VgR), resulting in the reduction in vitellogenin (Vg) deposition in oocytes. BmKr-h1 specifically bound the Kr-h1 binding site (KBS) between − 2818 and − 2805 nt upstream of BmVgR and enhanced the transcription of BmVgR. A 20E cis-regulatory element (CRE) was identified in the promoter of BmKr-h1 and functionally verified using luciferase reporter assay, EMSA and DNA-ChIP. Using pull-down assays, we identified a novel transcription factor B. mori Kr-h1 regulatory protein (BmKRP) that specifically bound the BmKr-h1 CRE and activated its transcription. CRISPR/Cas9-mediated knockout of BmKRP in female pupae suppressed the transcription of BmKr-h1 and BmVgR, resulting in arrested oogenesis. We identified BmKRP as a new transcription factor mediating 20E regulation of B. mori oogenesis. Our data suggests that induction of BmKRP by 20E regulates BmKr-h1 expression, which in turn induces BmVgR expression to facilitate Vg uptake and oogenesis. The online version contains supplementary material available at 10.1186/s12915-021-00952-2.
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