C/EBPβ and YY1 bind and interact with Smad3 to modulate lipopolysaccharide-induced amelotin gene transcription in mouse gingival epithelial cells
C/EBPβ and YY1 bind and interact with Smad3 to modulate lipopolysaccharide-induced amelotin gene transcription in mouse gingival epithelial cells
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C/EBPβ和YY1与Smad3结合并相互作用，调节小鼠牙龈上皮细胞中脂多糖诱导的amelotin基因转录
DOI：
10.1002/2211-5463.12566
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发表时间：
2018
期刊：
影响因子：
2.6
通讯作者：
Ogata Yorimasa
中科院分区：
文献类型：
--
作者：
Nakayama Yohei;Kobayashi Ryoki;Iwai Yasunobu;Noda Keisuke;Yamazaki Mizuho;Kurita-Ochiai Tomoko;Yoshimura Atsutoshi;Ganss Bernhard;Ogata Yorimasa
接合上皮（JE）在牙齿形成过程中由减少的牙釉质上皮发育而来，通过确保适当的免疫反应和牙龈上皮细胞的快速更新，对维持健康的牙周组织至关重要。我们之前已经证明炎症细胞因子与乙脑特异性基因（如AMTN）在牙龈上皮细胞中的表达之间存在关系。在此，我们阐明了牙龈卟啉单胞菌衍生的脂多糖（PgLPS）对amtngene转录的影响以及转录因子的相互作用。为了确定amtngene byPgLPS转录调控的分子基础，我们在小鼠牙龈上皮GE1细胞中使用与荧光素酶报告基因连接的小鼠seamtngene启动子进行了实时PCR和荧光素酶检测。凝胶迁移转移和染色质免疫沉淀试验用于鉴定与LPS诱导的转录因子结合的应答元件。接下来，我们通过免疫印迹和免疫沉淀分析了LPS诱导的转录因子的蛋白水平和转录因子之间的相互作用。LPS增加了amtnmrna水平，并提高了含有mouseAmtngene启动子- 116和- 238之间区域的构建体的荧光素酶活性。CCAAT/增强子结合蛋白（C/EBP） 1 -、C/EBP2 -和盈阳1 (YY1) -核蛋白复合物在LPS处理下增加。此外，我们还发现了LPS介导的与C/EBPβ、YY1和Smad3的相互作用。这些结果表明，pglps通过将C/ EBPβ-Smad3和YY1 - smad3复合物结合到mouseAmtngene启动子中的C/EBP1、C/EBP2和YY1应答元件上，调控amtngene的转录。
Junctional epithelium (JE) develops from reduced enamel epithelium during tooth formation and is critical for the maintenance of healthy periodontal tissue through ensuring appropriate immune responses and the rapid turnover of gingival epithelial cells. We have previously shown a relationship between inflammatory cytokines and expression of JE‐specific genes, such as amelotin (AMTN), in gingival epithelial cells. Here, we elucidated the effects ofPorphyromonas gingivalis‐derived lipopolysaccharide (PgLPS) onAmtngene transcription and the interaction of transcription factors. To determine the molecular basis of transcriptional regulation of theAmtngene byPgLPS, we performed real‐time PCR and carried out luciferase assays using a mouseAmtngene promoter linked to a luciferase reporter gene in mouse gingival epithelial GE1 cells. Gel mobility shift and chromatin immunoprecipitation assays were performed to identify response elements bound to LPS‐induced transcription factors. Next, we analyzed protein levels of the LPS‐induced transcription factors and the interaction of transcription factors by western blotting and immunoprecipitation. LPS increasedAmtnmRNA levels and elevated luciferase activities of constructs containing regions between −116 and −238 of the mouseAmtngene promoter.CCAAT/enhancer‐binding protein (C/EBP) 1–,C/EBP2– and Ying Yang 1 (YY1)–nuclear protein complexes were increased by LPS treatment. Furthermore, we identified LPS‐modulated interactions with C/EBPβ, YY1 and Smad3. These results demonstrate thatPgLPS regulatesAmtngene transcription via binding of C/EBPβ–Smad3 and YY1–Smad3 complexes to C/EBP1, C/EBP2 and YY1 response elements in the mouseAmtngene promoter.