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首页> 外文期刊>Molecular biology reports >Transcriptional activation of p21(WAF1/CIP1) is mediated by increased DNA binding activity and increased interaction between p53 and Sp1 via phosphorylation during replicative senescence of human embryonic fibroblasts
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Transcriptional activation of p21(WAF1/CIP1) is mediated by increased DNA binding activity and increased interaction between p53 and Sp1 via phosphorylation during replicative senescence of human embryonic fibroblasts

机译:p21(WAF1 / CIP1)的转录激活是通过人类胚胎成纤维细胞复制衰老过程中的DNA结合活性增强和p53与Sp1之间的相互作用通过磷酸化介导

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摘要

Although p21(WAF1/CIP1) is known to be elevated during replicative senescence of human embryonic fibroblasts (HEFs), the mechanism for p21 up-regulation has not been elucidated clearly. In order to explore the mechanism, we analyzed expression of p21 mRNA and protein and luciferase activity of full-length p21 promoter. The result demonstrated that p21 up-regulation was accomplished largely at transcription level. The promoter assay using serially-deleted p21 promoter constructs revealed that p53 binding site was the most important site and Sp1 binding sites were necessary but not sufficient for transcriptional activation of p21. In addition, p53 protein was shown to interact with Sp1 protein. The interaction was increased in aged fibroblasts and was regulated by phosphorylation of p53 and Sp1. DNA binding activity of p53 was significantly elevated in aged fibroblasts but that of Sp1 was not. DNA binding activities of p53 and Sp1 were also regulated by phosphorylation. Phosphorylation of p53 at serine-15 and of Sp1 at serines appears to be involved. Taken together, the result demonstrated that p21 transcription during replicative senescence of HEFs is up-regulated by increase in DNA binding activity and interaction between p53 and Sp1 via phosphorylation.
机译:尽管已知p21(WAF1 / CIP1)在人类胚胎成纤维细胞(HEFs)复制衰老过程中升高,但尚未清楚阐明p21上调的机制。为了探讨其机理,我们分析了全长p21启动子的p21 mRNA和蛋白表达以及萤光素酶活性。结果表明p21上调主要在转录水平上完成。使用序列缺失的p21启动子构建体的启动子分析表明,p53结合位点是最重要的位点,而Sp1结合位点对于p21的转录激活是必需的,但不足。另外,显示p53蛋白与Sp1蛋白相互作用。相互作用在老年成纤维细胞中增加,并受p53和Sp1磷酸化的调节。在老化的成纤维细胞中,p53的DNA结合活性显着提高,而Sp1的DNA结合活性则没有。 p53和Sp1的DNA结合活性也受到磷酸化作用的调节。似乎涉及丝氨酸15 p53和丝氨酸Sp1的磷酸化。两者合计,结果表明在HEFs的复制衰老过程中p21转录通过DNA结合活性的增加以及p53和Sp1之间通过磷酸化的相互作用而被上调。

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