Cooperative activation of Npr1 gene transcription and expression by interaction of Ets-1 and p300.
Cooperative activation of Npr1 gene transcription and expression by interaction of Ets-1 and p300.
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DOI：
10.1161/hypertensionaha.109.133033
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发表时间：
2009-07
期刊：
影响因子：
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通讯作者：
Pandey KN
中科院分区：
文献类型：
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作者：
Kumar P;Pandey KN
本研究旨在探讨Ets-1和p300在鸟苷酸环化酶-A/利钠肽受体-A（GC-A/NPRA）基因转录调控和表达中的协同作用。Ets-1和p300在小鼠系膜细胞（MMCs）中的过表达使Npr1启动子活性增加12倍，NPRA mRNA水平增加5倍，ANP依赖的cGMP细胞内积累增加26倍。用小干扰RNA（siRNA）敲低Ets-1和p300表达可抑制Npr1基因转录达90%。连续染色质免疫沉淀试验表明，P300和Ets-1之间的直接物理关联后，结合Npr1启动子，表明Ets-1和P300之间的物理相互作用是重要的，以增强Npr1基因转录。缺乏组蛋白乙酰转移酶（HAT）活性的突变体p300与Ets-1没有显示出功能效应，表明p300的HAT活性是调节Npr1基因转录的协同相互作用所必需的。野生型腺病毒E1A的过表达使Npr1启动子活性显著降低40%，而不能与p300结合的突变体E1A没有显示出任何影响。结果表明，Npr1基因的转录是由组蛋白乙酰转移酶p300和Ets-1的关键控制。目前的研究结果应该产生重要的见解的分子信号管理Npr1基因转录，在控制高血压和心血管事件的重要调节。
The objective of the present study was to gain insight into the cooperative role of Ets-1 and p300 in transcriptional regulation and expression of Npr1 gene (coding for guanylyl cyclase-A/natriuretic peptide receptor-A; GC-A/NPRA). Overexpression of Ets-1 and p300 in mouse mesangial cells (MMCs) increased Npr1 promoter activity by 12-fold, NPRA mRNA levels by 5-fold, and ANP-dependent intracellular accumulation of cGMP by 26-fold. Knockdown of Ets-1 and p300 expression by small interfering RNA (siRNA) inhibited Npr1 gene transcription by 90%. Sequential chromatin immunoprecipitation assay demonstrated a direct physical association between p300 and Ets-1 upon binding to Npr1 promoter, suggesting that a physical interaction between Ets-1 and p300 is important to enhance Npr1 gene transcription. Mutant p300 lacking histone acetyltransferase (HAT) activity did not show functional effect with Ets-1, suggesting that HAT activity of p300 is required for the cooperative interaction in modulating Npr1 gene transcription. Overexpression of wild-type adenovirus E1A significantly decreased the Npr1 promoter activity by 40%, whereas mutant E1A, which is incapable of binding to p300, did not show any effect. The results indicate that Npr1 gene transcription is critically controlled by histone acetyltransferase p300 and Ets-1. The present findings should yield important insights into the molecular signaling governing Npr1 gene transcription, an important regulator in the control of hypertension and cardiovascular events.