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HSF1 down-regulates XAF1 through transcriptional regulation   总被引:5,自引:0,他引:5  
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Porcine zygotic genome activation (ZGA) occurs along with global epigenetic remodeling at the 4-cell stage. These processes are regulated by histone acetylation, which requires acetyl-coenzyme A (CoA). Pyruvate dehydrogenase complex (PDC) is a crucial enzyme in glucose metabolism that converts pyruvate into acetyl-CoA. In mammalian cells, acetyl-CoA is produced by pyruvate dehydrogenase alpha 1 (PDHA1) translocated into the nucleus in special conditions. To determine whether zygotic PDHA1 plays a critical role in promoting histone acetylation during ZGA, a CRISPR/Cas9 genome editing system using multiple guide RNAs was employed to generate a PDHA1-targeted parthenogenetic embryo model. Results of immunofluorescent staining showed that the nuclear accumulation of PDHA1 during ZGA was significantly inhibited by PDHA1 targeting. Meanwhile, the 4-cell arrest rate significantly increased at 72 h after activation, indicating impeded embryonic development. In addition, nuclear histone acetylation significantly decreased when PDHA1 was targeted, and quantitative PCR showed that expression of several zygotic genes was significantly decreased in the PDHA1-targeting group compared to the control group. Overexpression of PDHA1 recovered the nuclear PDHA1, H3K9Ac and H3K27Ac and EIF1A expression levels. Moreover, the 5-to-8-cell-stage embryo development rate was only partially rescued. In conclusion, expression of zygotic origin PDHA1 contributes to porcine ZGA by maintaining histone acetylation in porcine embryos.  相似文献   

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目的:探讨烧伤血清诱导下热休克转录因子1(HSF1)对高迁移率族蛋白B1(HMGB1)的转录调控作用及其机制.方法:构建热休克转录因子1真核表达载体pcDNA3.1-HSF1,HMGB1野生型启动子荧光素酶报告基因pGL3-HMGB1-Y,HMGB1突变型启动子荧光素酶报告基因pGL3-HMGB1-T.pcDNA3.1-HSF1转染巨噬细胞RAW264.7,烧伤血清诱导后,半定量RT-PCR检测HMGB1 mRNA的表达.pcDNA3.1-HSF1,HMGB1启动子荧光素酶报告基因共转染RAW264.7,烧伤血清诱导后,检测并比较pGL3-HMGB1-Y和 pGL3-HMGB1-T的相对萤光素酶活性.结果:烧伤血清诱导下,HSF1可以下调RAW264.7 HMGB1mRNA的表达.pGL3-HMGB1-T的相对荧光素酶值较pGL3-HMGB1-Y明显下降,P<0.01,差异有显著统计学意义.结论:烧伤血清诱导下,HSF1可能通过与HMGB1启动子区HSE的结合下调小鼠巨噬细胞RAW264.7 HMGB1mRNA的表达.  相似文献   

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