Phosphorylation Regulates Functions of ZEB1 Transcription Factor.
Phosphorylation Regulates Functions of ZEB1 Transcription Factor.
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DOI：
10.1002/jcp.25338
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发表时间：
2016-10
影响因子：
5.6
通讯作者：
Cabanillas AM
中科院分区：
文献类型：
--
作者：
Llorens MC;Lorenzatti G;Cavallo NL;Vaglienti MV;Perrone AP;Carenbauer AL;Darling DS;Cabanillas AM
ZEB1转录因子在发育和疾病中都很重要，包括许多tgf β诱导的反应，以及许多肿瘤发生转移的上皮-间质转化（EMT）。ZEB1在不同的细胞类型中被不同的磷酸化；然而，磷酸化在ZEB1活性中的作用尚不清楚。荧光素酶报告基因研究和电泳迁移迁移试验（EMSA）表明，ZEB1磷酸化水平的降低增加了ZEB1靶基因的dna结合和转录抑制。对第二锌指结构域附近的ZEB1磷酸化位点突变体（称为ZD2）的功能分析表明，磷酸化的增加（由于PMA加离子霉素或IGF-1）可以抑制ZEB1-ZD2结构域克隆或全长ZEB1的转录抑制。该方法鉴定出对ZEB1的转录和dna结合活性有实质性调节作用的磷酸化位点。抗ZEB1抗体的免疫沉淀和磷酸化苏氨酸-脯氨酸特异性抗体的western分析表明，ERK共识位点Thr-867在ZEB1中被磷酸化。除了破坏EMSA测量的体外dna结合外，igf -1诱导的MEK/ERK磷酸化足以破坏GFP-ZEB1融合克隆的核定位。这些数据表明，ZEB1的磷酸化将tgf - β信号通路与其他信号通路（如IGF-1）整合在一起。这篇文章受版权保护。版权所有。
ZEB1 transcription factor is important in both development and disease, including many TGFβ-induced responses, and the epithelial-to-mesenchymal transition (EMT) by which many tumors undergo metastasis. ZEB1 is differentially phosphorylated in different cell types; however the role of phosphorylation in ZEB1 activity is unknown. Luciferase reporter studies and electrophoresis mobility shift assays (EMSA) show that a decrease in phosphorylation of ZEB1 increases both DNA-binding and transcriptional repression of ZEB1 target genes. Functional analysis of ZEB1 phosphorylation site mutants near the second zinc finger domain (termed ZD2) show that increased phosphorylation (due to either PMA plus ionomycin, or IGF-1) can inhibit transcriptional repression by either a ZEB1-ZD2 domain clone, or full-length ZEB1. This approach identifies phosphosites that have a substantial effect regulating the transcriptional and DNA-binding activity of ZEB1. Immunoprecipitation with anti-ZEB1 antibodies followed by western analysis with a phospho-Threonine-Proline-specific antibody indicates that the ERK consensus site at Thr-867 is phosphorylated in ZEB1. In addition to disrupting in vitro DNA-binding measured by EMSA, IGF-1-induced MEK/ERK phosphorylation is sufficient to disrupt nuclear localization of GFP-ZEB1 fusion clones. These data suggest that phosphorylation of ZEB1 integrates TGFβ signaling with other signaling pathways such as IGF-1. This article is protected by copyright. All rights reserved.
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发表时间：
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