Identification of prohibitin and prohibiton as novel factors binding to the p53 induced gene 3 (PIG3) promoter (TGYCC)(15) motif.
Identification of prohibitin and prohibiton as novel factors binding to the p53 induced gene 3 (PIG3) promoter (TGYCC)(15) motif.
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DOI：
10.1016/j.bbrc.2013.12.124
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发表时间：
2014-01-24
影响因子：
3.1
通讯作者：
Wei, Qingyi
中科院分区：
文献类型：
--
作者：
Guan, Xiaoxiang;Liu, Zhensheng;Wang, Luo;Johnson, David G.;Wei, Qingyi
p53诱导基因3（PIG 3）的启动子含有被称为p53结合位点的五核苷酸（TGYCC）n的可变数目的串联重复（VNTR）。在这项研究中，我们研究了其他潜在的分子是否可以结合到这个PIG 3启动子（TGYCC）n基序。配体色谱结合液相色谱-串联质谱分析表明，直接相互作用的prohibitin和/或arabiton与（TGYCC）15基序，这是证实了电泳迁移率变动分析和超凝胶位移分析与抗prohibitin和抗arabiton抗体。使用染色质免疫沉淀分析，我们进一步证明，无论p53状态和凋亡应激如何，抑制素和抑制蛋白在体内与（TGYCC）15基序相关。我们还发现，prohibitin和arabiton上调PIG 3的转录不依赖于p53，虽然p53明显增强了这一过程，并且，敲低prohibitin和arabiton抑制喜树碱诱导的细胞凋亡。总之，我们的研究结果表明，抑制素和抑制素通过与PIG 3启动子（TGYCC）15基序结合而促进PIG 3介导的细胞凋亡。
The promoter of p53 induced gene 3 (PIG3) contains a variable number of tandem repeats (VNTRs) of pentanucleotides (TGYCC)n that is known as a p53 binding site. In this study, we investigated whether other potential molecules could bind to this PIG3 promoter (TGYCC)n motif. Ligand-chromatography combined with liquid chromatography–tandem mass spectrometry analyses indicated direct interactions of prohibitin and/or prohibiton with the (TGYCC)15 motif, which was confirmed by electrophoretic mobility shift assay and super-gel shift analysis with anti-prohibitin and anti-prohibiton antibodies. Using the chromatin immunopercipipation assay, we further demonstrated that prohibitin and prohibiton associated with the (TGYCC)15 motif in vivo regardless of the p53 status and apoptotic stress. We also found that prohibitin and prohibiton up-regulated PIG3 transcription independent of p53, although p53 obviously enhanced this process, and that the knock-down of prohibitin and prohibiton inhibited camptothecin-induced apoptosis. Taken together, our findings suggest that prohibitin and prohibiton contribute to PIG3-mediated apoptosis by binding to the PIG3 promoter (TGYCC)15 motif.
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