Promoter elements and transcriptional control of the chicken β tropomycin gene
Promoter elements and transcriptional control of the chicken β tropomycin gene
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鸡β原霉素基因的启动子元件和转录控制
DOI：
10.1093/nar/22.10.1838
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发表时间：
1994
期刊：
影响因子：
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通讯作者：
M. Lemonnier
中科院分区：
文献类型：
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作者：
M. Toutant;C. Gauthier;M. Fiszman;M. Lemonnier
鸡β -原肌球蛋白（β - TM）基因有两个转录起始位点（sk位点和nmCAP位点），分别用于肌肉组织和非肌肉组织。为了了解β TM基因的组织特异性和发育调控表达的机制，我们分析了与每个CAP位点相关的5'区。在细菌氯霉素乙酰转移酶（chloramphenicol acetyltransferase， CAT）报告基因上游插入nmCAP位点截断的5'区域，并将这些构建物转染到禽的肌源性和非肌源性细胞中。在所有细胞类型中，最大转录是由CAT结构（-168/ + 216 nt）驱动的。先前对β TMskCAP位点的区域5'的缺失分析表明，805nt赋予肌管特异性转录。在这项工作中，我们表征了一个增强子元件（-201/-68 nt），它包含一个E盒（-177），一个变体CArG盒（-104）和一个7Cs延伸（-147）。任何这些基序的突变都会导致肌管特异性转录活性的降低。电泳迁移位移测定表明，这些顺式作用序列特异性结合核蛋白。这个增强器以方向依赖的方式起作用。
The chicken beta tropomyosin (beta TM) gene has two alternative transcription start sites (sk and nmCAP sites) which are used in muscle or non muscle tissues respectively. In order to understand the mechanisms involved in the tissue-specific and developmentally-regulated expression of the beta TM gene, we have analyzed the 5' regions associated with each CAP site. Truncated regions 5' to the nmCAP site were inserted upstream to the bacterial chloramphenicol acetyltransferase (CAT) reporter gene and these constructs were transfected into avian myogenic and non myogenic cells. The maximum transcription is driven by the CAT construct (-168/ + 216 nt) in all cell types. Previous deletion analysis of the region 5' to the beta TMskCAP site has indicated that 805 nt confer myotube-specific transcription. In this work, we characterized an enhancer element (-201/-68 nt) which contains an E box (-177), a variant CArG box (-104) and a stretch of 7Cs (-147). Mutation of any of these motifs results in a decrease of the myotube-specific transcriptional activity. Electrophoretic mobility shift assays indicate that these cis-acting sequences specifically bind nuclear proteins. This enhancer functions in an orientation-dependent manner.