ANKRD1 Acts as a Transcriptional Repressor of MMP13 via the AP-1 Site
ANKRD1 Acts as a Transcriptional Repressor of MMP13 via the AP-1 Site
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DOI：
10.1128/mcb.01357-13
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发表时间：
2014-04-01
影响因子：
5.3
通讯作者：
Davidson, Jeffrey M.
中科院分区：
文献类型：
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作者：
Almodovar-Garcia, Karinna;Kwon, Minjae;Davidson, Jeffrey M.
转录辅因子ANKRD 1在创伤修复过程中被急剧诱导，并且其过表达促进愈合。我们最近发现，小鼠Ankrd 1的整体缺失损害伤口收缩，并增强缺血性伤口的坏死。Ankrd 1（-/-）（KO）成纤维细胞的定量PCR阵列表明，ANKRD 1调节MMP基因。酵母双杂交和免疫共沉淀分析将ANKRD 1与核仁素相关联，核仁素抑制MMP 13的AP-1活化。Ankrd 1缺失增强了基础和佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）诱导的MMP 13启动子活性;相反，Ankrd 1在对照细胞中的过表达降低了PMA诱导的MMP 13启动子活性。在KO成纤维细胞中Ankrd 1重建降低MMP 13 mRNA，而Ankrd 1敲除增加这些水平。KO与Ankrd 1（fl/fl）（FLOX）小鼠相比，MMP 13 mRNA和蛋白在完整皮肤和伤口中升高。电泳迁移率变动分析凝胶移动模式表明，在Ankrd 1不存在的情况下，额外的转录因子与MMP 13 AP-1位点结合，并且通过染色质免疫沉淀分析加强了这一概念，因为与KO成纤维细胞相比，FLOX提取物中c-Jun与AP-1位点的结合更强。我们认为ANKRD 1与核仁素等因子一起抑制MMP 13的转录。此外，Ankrd 1缺失还减轻了MMP 10的转录抑制。核ANKRD 1似乎通过MMPs调节细胞外基质重塑。
The transcriptional cofactor ANKRD1 is sharply induced during wound repair, and its overexpression enhances healing. We recently found that global deletion of murine Ankrd1 impairs wound contraction and enhances necrosis of ischemic wounds. A quantitative PCR array of Ankrd1(-/-) (KO) fibroblasts indicated that ANKRD1 regulates MMP genes. Yeast two-hybrid and co-immunoprecipitation analyses associated ANKRD1 with nucleolin, which represses AP-1 activation of MMP13. Ankrd1 deletion enhanced both basal and phorbol 12-myristate 13-acetate (PMA)-induced MMP13 promoter activity; conversely, Ankrd1 overexpression in control cells decreased PMA-induced MMP13 promoter activity. Ankrd1 reconstitution in KO fibroblasts decreased MMP13 mRNA, while Ankrd1 knockdown increased these levels. MMP13 mRNA and protein were elevated in intact skin and wounds of KO versus Ankrd1(fl/fl) (FLOX) mice. Electrophoretic mobility shift assay gel shift patterns suggested that additional transcription factors bind to the MMP13 AP-1 site in the absence of Ankrd1, and this concept was reinforced by chromatin immunoprecipitation analysis as greater binding of c-Jun to the AP-1 site in extracts from FLOX versus KO fibroblasts. We propose that ANKRD1, in association with factors such as nucleolin, represses MMP13 transcription. Ankrd1 deletion additionally relieved MMP10 transcriptional repression. Nuclear ANKRD1 appears to modulate extracellular matrix remodeling by MMPs.