YY1-binding sites provide central switch functions in the PARP-1 gene expression network.

YY1-binding sites provide central switch functions in the PARP-1 gene expression network.
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YY1结合位点在PARP-1基因表达网络中提供中央开关功能。

DOI:
10.1371/journal.pone.0044125
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发表时间:
2012
期刊:
影响因子:
3.7
通讯作者:
Vidaković M
Vidaković M
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Doetsch M;Gluch A;Poznanović G;Bode J;Vidaković M

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有证据表明转录因子 Yin Yang 1 (YY1) 和聚(ADP-核糖)聚合酶-1 (PARP-1) 之间的相互作用参与调节小鼠 PARP-1 基因 (muPARP-1) 启动子活性。我们在 muPARP-1 核心启动子 (−574/+200) 的七个位置上鉴定了潜在的 YY1 结合基序 (BM)。使用抗 YY1 抗体和带有核心启动子的线性化或超螺旋形式的质粒,以及含有单个 YY1 结合基序和四个 muPARP-1 启动子片段的 30 bp 寡核苷酸探针,通过电泳超位移测定观察 YY1 的结合。我们检测到 YY1 与 BM1 (−587/−558)、BM4 (−348/−319) 结合,并与 BM7 (+86/+115) 存在非常显着的关联。对 BM7 的检查揭示了 muPARP-1 翻译起始位点与 Kozak 序列以及 YY1 和 PARP-1 识别位点的重叠。 YY1 和 PARP-1 核心基序的定点诱变消除了蛋白质结合,并表明 YY1 介导与 Kozak 序列相邻的 PARP-1 结合。在 muPARP-1 启动子控制下的报告基因转染实验表明,YY1 与 BM1 和 BM4 的结合独立地抑制了启动子。这些位点的突变阻止了 YY1 结合,从而增加了报告基因的活性。在 PARP-1 过表达的 PARP-1 敲除细胞中,类似于 YY1 的效果变得明显; YY1 和 PARP-1 的过度表达揭示了它们的协同作用。与我们之前的发现一起,这些结果通过 YY1 作用扩展了 PARP-1 自动调节环原理,这意味着 muPARP-1 表达的严格限制。 PARP-1 和 YY1 的联合作用对细胞稳态做出了重要贡献。
Evidence is presented for the involvement of the interplay between transcription factor Yin Yang 1 (YY1) and poly(ADP-ribose) polymerase-1 (PARP-1) in the regulation of mouse PARP-1 gene (muPARP-1) promoter activity. We identified potential YY1 binding motifs (BM) at seven positions in the muPARP-1 core-promoter (−574/+200). Binding of YY1 was observed by the electrophoretic supershift assay using anti-YY1 antibody and linearized or supercoiled forms of plasmids bearing the core promoter, as well as with 30 bp oligonucleotide probes containing the individual YY1 binding motifs and four muPARP-1 promoter fragments. We detected YY1 binding to BM1 (−587/−558), BM4 (−348/−319) and a very prominent association with BM7 (+86/+115). Inspection of BM7 reveals overlap of the muPARP-1 translation start site with the Kozak sequence and YY1 and PARP-1 recognition sites. Site-directed mutagenesis of the YY1 and PARP-1 core motifs eliminated protein binding and showed that YY1 mediates PARP-1 binding next to the Kozak sequence. Transfection experiments with a reporter gene under the control of the muPARP-1 promoter revealed that YY1 binding to BM1 and BM4 independently repressed the promoter. Mutations at these sites prevented YY1 binding, allowing for increased reporter gene activity. In PARP-1 knockout cells subjected to PARP-1 overexpression, effects similar to YY1 became apparent; over expression of YY1 and PARP-1 revealed their synergistic action. Together with our previous findings these results expand the PARP-1 autoregulatory loop principle by YY1 actions, implying rigid limitation of muPARP-1 expression. The joint actions of PARP-1 and YY1 emerge as important contributions to cell homeostasis.
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发表时间: 2010-12-22
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发表时间: 2009-11-01
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DOI: 10.1160/th04-09-0605
发表时间: 2005-02-01
影响因子: 6.7
作者:
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DOI: 10.1016/0921-8777(89)90012-8
发表时间: 1989-09-01
期刊: MUTATION RESEARCH
影响因子: --
作者:
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