ZNF300 tight self-regulation and functioning through DNA methylation and histone acetylation.
ZNF300 tight self-regulation and functioning through DNA methylation and histone acetylation.
复制标题
ZNF300 通过 DNA 甲基化和组蛋白乙酰化进行严格的自我调节和发挥作用
DOI：
10.1186/s13578-017-0160-8
复制
发表时间：
2017
影响因子：
7.5
通讯作者：
Zhang JJ
中科院分区：
文献类型：
--
作者：
Yan FJ;Fan J;Huang Z;Zhang JJ
背景越来越多的证据表明KRAB-ZNFs参与多种生物过程。作为KRAB-ZNFs的典型成员，ZNF300的失调有助于多种病理，如白血病和癌症。然而，ZNF 300的紧密调控及其病理生理功能的机制仍然在很大程度上unknown. MethodsZNF 300 ZFR对基因转录活性的影响进行了测量双荧光素酶报告系统。ChIP-PCR检测ZNF300蛋白和H3K9Ac在ZNF300基因中的富集。采用免疫共沉淀法和Western blot检测ZNF 300与KAP 1的相互作用。采用BSP分析ZNF 300基因启动子DNA甲基化。结果在本研究中，我们发现ZNF 300的锌指结构域编码区（ZFR）可能通过介导DNA甲基化而发挥阻遏物的作用，并且ZNF 300与ZNF 300的ZFR结合，提示ZNF 300可能具有自抑制机制。为了支持这一点，DNA甲基化抑制上调ZNF300表达，ZNF300过表达抑制内源性ZNF300表达。更重要的是，DNA甲基化抑制恢复了ZNF300下调抑制的K562细胞中的巨核细胞分化，表明DNA甲基化在ZNF300功能中的重要作用。有趣的是，ZNF300敲低恢复全球H3K9Ac减少K562细胞经历megakaryocyte differentiation.ConclusionsOur研究揭示了新的功能ZNF300，可能介导其调节和功能，通过调节表观遗传修饰。
BackgroundAccumulating evidence demonstrates that the KRAB-ZNFs involve in various biological processes. As a typical member of KRAB-ZNFs, dysregulation of ZNF300 contributes to multiple pathologies such as leukemia and cancer. However, mechanisms underlying ZNF300 tight regulation and its pathophysiological function remain largely unknown.MethodsThe effect of ZNF300ZFR on gene transcriptional activity was measured by Dual luciferase reporter system. ChIP-PCR assay were performed to detect the enrichment of ZNF300 protein and H3K9Ac in the ZNF300 gene. Co-immunoprecipitation assays followed by western blot were performed to detect the interaction between ZNF300 and KAP1. The DNA methylation in the ZNF300 gene promoter was analyzed by BSP. ZNF300 function on K562 cell differentiation was analyzed by flow cytometry.ResultsIn this study, we found that the zinc finger domain-encoding region (ZFR) of ZNF300 functioned as a repressor possibly by mediating DNA methylation and ZNF300 bound to its ZNF300ZFR, suggesting a potential auto-inhibition mechanism. To support this, DNA methylation inhibition upregulated ZNF300 expression and ZNF300 overexpression inhibited endogenous ZNF300 expression. More importantly, DNA methylation inhibition restored megakaryocyte differentiation in K562 cells suppressed by ZNF300 downregulation, suggesting an important role of DNA methylation in ZNF300 function. Interestingly, ZNF300 knockdown restored global H3K9Ac that was reduced in K562 cells undergoing megakaryocyte differentiation.ConclusionsOur study revealed novel features of ZNF300 that possibly mediate its regulation and function by modulating epigenetic modifications.