Interacting C/EBPg and YBP regulate DNA methyltransferase 1 expression in Bombyx mori embryos and ovaries
Interacting C/EBPg and YBP regulate DNA methyltransferase 1 expression in Bombyx mori embryos and ovaries
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相互作用的 C/EBPg 和 YBP 调节家蚕胚胎和卵巢中 DNA 甲基转移酶 1 的表达
DOI：
10.1016/j.ibmb.2021.103583
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发表时间：
2021-05-18
影响因子：
3.8
通讯作者：
Zheng, Sichun
中科院分区：
文献类型：
--
作者：
Lyu, Hao;Xu, Guanfeng;Zheng, Sichun
DNA甲基化是一种重要的表观遗传修饰。DNA甲基转移酶（Dnmts）催化5-甲基胞嘧啶的形成，在一些昆虫的卵巢和胚胎发育中起重要作用。然而，Dnmt介导卵巢和胚胎发育的潜在机制尚不清楚。本研究对家蚕Dnmt1的调控及其功能进行了研究。通过逐步删除Dnmt1上游的序列，发现位于转录起始位点-580和-560区域之间的区域具有最高的转录活性。电泳迁移率转移试验和染色质免疫沉淀表明，转录因子Y盒结合蛋白（YBP）是人类Y盒结合蛋白1 （YBX1）的同源物，结合在-580到-560区域。在家蚕细胞系中，YBP敲低和过表达表明YBP激活Dnmt1的表达。此外，gst -pull - down和共免疫沉淀表明YBP与卵巢CCAAT/增强子结合蛋白（C/EBPg）可以相互结合。同时敲除C/EBPg和YBP在抑制Dnmt1表达和降低孵化率方面比单基因RNAi更有效。这些结果表明，C/ EBPg和YBP的相互作用激活了Dnmt1的表达。结合研究基因的表达谱，我们的研究结果表明，C/EBPg和YBP在卵巢和胚胎中的高水平表达和相互作用增强了Dnmt1的表达，从而保证了家蚕的高繁殖率。
DNA methylation is an important epigenetic modification. DNA methyltransferases (Dnmts), which catalyze the formation of 5-methylcytosine, play a role in ovarian and embryonic development in some insects. However, the underlying mechanism of Dnmt in mediating ovarian and embryonic development remains unclear. In this study, the regulation and function of Bombyx mori Dnmt1 were investigated. By progressively deleting the sequence upstream of Dnmt1, a region located between -580 and -560 region from the transcription initiation site was found to have the most transcriptional activity. Electrophoretic mobility shift assay and chromatin immunoprecipitation demonstrated that transcription factor Y box binding protein (YBP), a homolog of human Y box binding protein 1 (YBX1), bound to the -580 to -560 region. YBP knockdown and overexpression in a Bombyx cell line indicated that YBP activates Dnmt1 expression. Furthermore, GST-pulldown and coimmunoprecipitation demonstrated that YBP and ovarian CCAAT/enhancer binding protein (C/EBPg) could bind each other. Simultaneous knockdown of C/EBPg and YBP was more effective than single-gene RNAi in inhibiting Dnmt1 expression and reducing the hatching rate. These results demonstrated that the interaction of C/ EBPg and YBP activated Dnmt1 expression. Correlated with the expression profiles of the studies genes, our results suggest that high-level expression and interaction of C/EBPg and YBP in ovaries and embryos enhance the expression of Dnmt1, thus ensuring high reproduction rate in B. mori.