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Dynamic and distinct histone modifications modulate the expression of key adipogenesis regulatory genes

机译:动态和独特的组蛋白修饰可调节关键脂肪形成调控基因的表达

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Histone modifications and their modifying enzymes are fundamentally involved in the epigenetic regulation of adipogenesis. This study aimed to define the roles of various histone modifications and their "division of labor" in fat cell differentiation. To achieve these goals, we examined the distribution patterns of eight core histone modifications at five key adipogenic regulatory genes, Pref-1, C/EBPβ, C/EBPα, PPARγ2 and aP2, during the adipogenesis of C3H 10T1/2 mouse mesenchymal stem cells (MSCs) and 3T3-L1 preadipocytes. We found that the examined histone modifications are globally stable throughout adipogenesis but show distinct and highly dynamic distribution patterns at specific genes. For example, the Pref-1 gene has lower levels of active chromatin markers and significantly higher H3 K27 tri-methylation in MSCs compared with committed preadipocytes; the C/EBPβ gene is enriched in active chromatin markers at its 3?-UTR; the C/EBPα gene is predominantly marked by H3 K27 tri-methylation in adipogenic precursor cells, and this repressive marker decreases dramatically upon induction; the PPARγ2 and aP2 genes show increased histone acetylation on both H3 and H4 tails during adipogenesis. Further functional studies revealed that the decreased level of H3 K27 tri-methylation leads to de-repression of Pref-1 gene, while the increased level of histone acetylation activates the transcription of PPARγ2 and aP2 genes. Moreover, the active histone modification-marked 3′-UTR of C/EBPβ gene was demonstrated as a strong enhancer element by luciferase assay. Our results indicate that histone modifications are gene-specific at adipogenic regulator genes, and they play distinct roles in regulating the transcriptional network during adipogenesis.
机译:组蛋白修饰及其修饰酶从根本上参与脂肪形成的表观遗传调控。这项研究旨在确定各种组蛋白修饰及其在脂肪细胞分化中的“分工”作用。为了实现这些目标,我们检查了C3H 10T1 / 2小鼠间充质干细胞成脂过程中五个主要成脂调控基因Pref-1,C /EBPβ,C /EBPα,PPARγ2和aP2的八个核心组蛋白修饰的分布模式。 (MSC)和3T3-L1前脂肪细胞。我们发现所检查的组蛋白修饰在整个成脂过程中是全局稳定的,但在特定基因上显示出独特且高度动态的分布模式。例如,与定型前脂肪细胞相比,MSC中的Pref-1基因具有较低的活性染色质标记水平和显着较高的H3 K27三甲基化水平。 C /EBPβ基因在其3′-UTR处富含活性染色质标记。 C /EBPα基因主要由成脂前体细胞中的H3 K27三甲基化标记,并且该诱导性标记在诱导后急剧下降。在脂肪形成过程中,PPARγ2和aP2基因在H3和H4尾巴上均显示出组蛋白乙酰化增加。进一步的功能研究表明,H3 K27三甲基化水平的降低导致Pref-1基因的抑制,而组蛋白乙酰化水平的提高则激活了PPARγ2和aP2基因的转录。此外,通过荧光素酶测定法证实了C /EBPβ基因的活性组蛋白修饰标记的3'-UTR是强的增强子元件。我们的结果表明,组蛋白修饰在成脂调节基因上是基因特异性的,在成脂过程中它们在调节转录网络中起着独特的作用。

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