Myocyte enhancer factor 2 regulates expression of medaka Oryzias latipes fast skeletal myosin heavy chain genes in a temperature-dependent manner
Myocyte enhancer factor 2 regulates expression of medaka Oryzias latipes fast skeletal myosin heavy chain genes in a temperature-dependent manner
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DOI：
10.1016/j.gene.2007.09.016
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发表时间：
2008-01-15
期刊：
影响因子：
3.5
通讯作者：
Watabe, Shugo
中科院分区：
文献类型：
--
作者：
Liang, Chun-Shi;Ikeda, Daisuke;Watabe, Shugo
我们的特点是快速骨骼肌球蛋白重链基因（MYH）的启动子活性青鳉。在青鳉MYH、mMYH 10和inMY 1130中，分别主要在适应于10 ℃和30 ℃的青鳉中表达的类似于6 kb的5-侧翼区含有各种顺式元件，这些顺式元件被认为与转录调节因子如MyoD和肌细胞增强因子2（MEF 2）家族成员以及活化的T细胞的核因子结合。为了以温度依赖性方式定位负责mMYH表达的功能区域，将从5 '侧翼区域制备的一系列缺失和位点突变构建体融合到市售质粒中的荧光素酶基因上，并直接注射到适应10 ℃和30 ℃的青鳉背侧快肌中。MEF 2结合位点位于mMYH 10的5 '侧翼区的-966至-957处的截短导致在10 ℃下的不同基因表达。通过突变构建体进一步证实了通过去除该结合位点的活化效果。MyoD家族成员被认为与之结合的E盒位点之一位于mMYH 10的-613至-607处，并被发现负责转录活性。相反，位于mMYH 30的-960至-951的MEF 2结合位点参与30 ℃下的活化。因此，这些瞬时转染试验证明MEF 2结合位点对于mMYH的温度依赖性表达至关重要。(c)2007 Elsevier B. V.保留所有权利。
We characterized the promoter activity of fast skeletal myosin heavy chain genes (MYHs) from medaka Oryzias latipes. The 5-flanking region of similar to 6 kb in medaka MYHs, mMYH10 and inMY1130, predominantly expressed in medaka acclimated to 10 degrees C and 30 degrees C, respectively, contained various cis-elements that are supposed to bind to transcriptional regulatory factors such as MyoD) and myocyte enhancer factor 2 (MEF2) family members and nuclear factor of activated T cells. To localize functional regions responsible for the mMYH expression in a temperature-dependent manner, a series of deletion and site mutation constructs prepared from the 5'-flanking regions were fused to the luciferase gene in a commercially available plasmid and directly injected into the dorsal fast muscle of medaka acclimated to 10 degrees C and 30 degrees C. The truncation of MEF2 binding site located at -966 to -957 in the 5'-flanking region of mMYH10 resulted in distinct gene expression at 10 degrees C. The activation effect by the removal of this binding site was further confirmed by the mutation construct. One of the E box sites, to which MyoD family members are supposed to bind, was located at -613 to -607 of mMYH10, and found to be responsible for the transcriptional activity. In contrast, the MEF2 binding site located at -960 to -951 of mMYH30 was involved in the activation at 30 degrees C. Thus, these transient transfection assays demonstrated that the MEF2 binding site is crucial for a temperature-dependent expression of mMYHs. (c) 2007 Elsevier B.V. All rights reserved.