Homeobox protein A1-like (Hox A1-like) and DNA methylation regulate embryo-specific ZnF 615 gene expression and embryonic development in the silkworm Bombyx mori
Homeobox protein A1-like (Hox A1-like) and DNA methylation regulate embryo-specific ZnF 615 gene expression and embryonic development in the silkworm Bombyx mori
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同源盒蛋白 A1 样（Hox A1 样）和 DNA 甲基化调节家蚕胚胎特异性 ZnF 615 基因表达和胚胎发育
DOI：
10.1111/1744-7917.13152
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通讯作者：
Guan-Feng Xu
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作者：
Yu-Lin Tian;Tong-Yu Fu;Qi-En Zhong;Yi-Guang Lin;Si-Chun Zheng;Guan-Feng Xu
DNA 甲基化和转录因子在基因表达和动物发育中发挥作用。在昆虫中，DNA 甲基化修饰基因体，但 DNA 甲基化和转录因子如何调节基因表达尚不清楚。在本研究中，我们研究了家蚕锌指蛋白615（ZnF 615）（DNA甲基转移酶1（Dnmt1）下游基因）表达的调节机制及其对胚胎发育调控的影响。通过逐步截断ZnF 615启动子，发现含有同源盒（Hox）蛋白顺式调控元件（CRE）的-223和-190nt区域对ZnF 615的转录影响最大。RNA干扰（RNAi）介导的Hox家族基因的敲低和过表达表明Hox A1-like可以增强ZnF 615的信使RNA水平。进一步研究表明 Hox A1 样通过直接结合其启动子的 -223 和 -190 nt 区域来调节 ZnF 615 的表达。同时 RNAi 介导的 Hox A1-like 和 Dnmt1 敲低或过表达显着抑制或增强任一基因单独对 ZnF 615 表达的调节作用，表明基因体的 DNA 甲基化和转录因子与启动子的结合对于基因表达至关重要。 RNAi介导的Hox A1-like和Dnmt1敲低表明胚胎发育迟缓，孵化率降低。综上所述，这些数据表明 Hox A1 样和 DNA 甲基化增强了 ZnF 615 的表达，从而影响 B.莫里胚胎。
DNA methylation and transcription factors play roles in gene expression and animal development. In insects, DNA methylation modifies gene bodies, but how DNA methylation and transcription factors regulate gene expression is unclear. In this study, we investigated the mechanism that regulates the expression ofBombyx moriZinc finger protein 615 (ZnF 615), which is a downstream gene of DNA methyltransferase 1 (Dnmt1), and its effects on the regulation of embryonic development. By progressively truncating theZnF 615promoter, it was found that the −223 and −190 nt region, which contains homeobox (Hox) proteincis‐regulatory elements (CREs), had the greatest impact on the transcription ofZnF 615. RNA interference (RNAi)‐mediated knockdown and overexpression ofHoxfamily genes showed that Hox A1‐like can enhance the messenger RNA level ofZnF 615. Further studies showed that Hox A1‐like regulatesZnF 615expression by directly binding to the −223 and −190 nt region of its promoter. Simultaneous RNAi‐mediated knockdown or overexpression ofHox A1‐likeandDnmt1significantly inhibited or enhanced the regulatory effect of either gene alone onZnF 615expression, suggesting that both DNA methylation of gene bodies and binding of transcription factors to promoters are essential for gene expression. RNAi‐mediated knockdown ofHox A1‐likeandDnmt1showed that the embryonic development was retarded and the hatching rate was decreased. Taken together, these data suggest that Hox A1‐like and DNA methylation enhance the expression ofZnF 615, thereby affecting the development ofB. moriembryos.