Interaction of Smads with collagen types I, III, and V.
Interaction of Smads with collagen types I, III, and V.
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Smads 与 I、III 和 V 型胶原蛋白的相互作用。
DOI：
10.1016/j.bbrc.2003.09.130
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发表时间：
2003
影响因子：
3.1
通讯作者：
Pisano,MMichele
中科院分区：
文献类型：
--
作者：
Ellis,LeslieR;Warner,DennisR;Greene,RobertM;Pisano,MMichele
哺乳动物口面部的个体发育受许多发育信号控制，包括由转化生长因子β（TGFβs）启动的信号。在小鼠中靶向缺失编码几种TGFβ的基因已被证明会导致继发性腭裂。生长因子的TGFβ家族的成员利用细胞内Smads作为信号转导子。Smads 2和3是转录调节因子，通过其保守的MH 1结构域结合DNA，并通过其MH 2结构域激活/抑制TGFβ应答基因的转录。使用酵母双杂交筛选从胎鼠orofacial组织构建的cDNA表达文库，我们已经确定了三种类型的胶原蛋白（I型，III型和V型），能够结合到MH 2结构域的Smad 3。这些相互作用证实了谷胱甘肽S-转移酶（GST）下拉分析，其中MH 2结构域的Smad 3融合GST强烈相互作用，在体外翻译，35 S标记的胶原蛋白I型，III型和V型。每种胶原蛋白也结合到MH 2结构域的Smads 4和7，并在较小程度上，全长Smads 1，2，3，和4。Smads与胶原蛋白的结合是一个新的观察结果。此外，TGFβ是哺乳动物口面发育过程中胶原合成和周转的有效调节剂。因此，这些数据表明了TGFβ信号级联反馈调节的重要手段。
Ontogenesis of the mammalian orofacial region is controlled by numerous developmental signals, including those initiated by the transforming growth factors β (TGFβs). Targeted deletion of the genes encoding several of the TGFβs in mice has been shown to result in clefts of the secondary palate. Members of the TGFβ family of growth factors utilize intracellular Smads as signal transducers. Smads 2 and 3 are transcriptional regulators that bind DNA through their conserved MH1 domains and activate/inhibit transcription of TGFβ-responsive genes through their MH2 domains. Using a yeast two-hybrid screen of a cDNA expression library constructed from fetal murine orofacial tissue, we have identified three types of collagens (types I, III, and V) that are capable of binding to the MH2 domain of Smad 3. These interactions were confirmed by glutathione S-transferase (GST) pull-down assays in which the MH2 domain of Smad 3 fused to GST interacted strongly with in vitro translated,35S-labeled collagen types I, III, and V. Each collagen also bound to the MH2 domains of Smads 4 and 7 and, to a lesser extent, full-length Smads 1, 2, 3, and 4. Binding of Smads to collagen is a novel observation. Moreover, TGFβ is a potent regulator of collagen synthesis and turnover during mammalian orofacial development. These data thus suggest an important means of feedback regulation of the TGFβ signaling cascade.