Boam D S, Davidson I, Chambon P
Institut de Génétique et de Biologie Moléculair et Cellulaire, Parc d'Innovations, Illkirch, Strasbourg, France.
J Biol Chem. 1995 Aug 18;270(33):19487-94. doi: 10.1074/jbc.270.33.19487.
TEF-1 is a tissue-specific human transcription factor which binds to and activates transcription from the SV40 early promoter and the HPV-16 E6/E7 promoter and may be involved in regulation of muscle-specific and placenta-specific gene expression. To investigate the mechanism of its tissue-specific expression, we have isolated up to 3 kilobase pairs of 5'-flanking DNA and characterized the promoter of the gene for TEF-1. Multiple transcription start sites centering on a motif similar to the initiator element (Inr) were identified. A minimal promoter, which contains no recognizable TATA element but contains an Inr, delimited at -137 base pairs had full transcriptional activity both in vivo in HeLa cells and in vitro in HeLa cell extracts. This promoter is also highly active in vitro in lymphoid cell extracts, but not in vivo in lymphoid cell lines, which do not express the endogenous TEF-1 gene. The minimal promoter, which is sufficient to direct tissue-specific expression of the TEF-1 gene in vivo, contains multiple sites which bind the ubiquitous transcription factors Sp1 and ATF-1. Mutation of the Inr completely abolished transcription from the major start site while transcription from the minor sites was slightly augmented. Inactivation of the proximal Sp1 site abolished transcription from the principle start site and increased transcription from a 5' minor start site. Insertion of a TATA box element did not qualitatively alter the pattern of start site usage which seemed to be dependent upon integrity of the upstream Sp1 site. These observations suggest a "cross-talk" between the Inr and a proximal element to fix transcription start sites, which is independent of spacing and the presence of a TATA element.
TEF-1是一种组织特异性的人类转录因子,它能结合并激活SV40早期启动子和HPV-16 E6/E7启动子的转录,可能参与肌肉特异性和胎盘特异性基因表达的调控。为了研究其组织特异性表达的机制,我们分离了长达3千碱基对的5'侧翼DNA,并对TEF-1基因的启动子进行了表征。确定了多个以类似于起始子元件(Inr)的基序为中心的转录起始位点。一个最小启动子,它不包含可识别的TATA元件但包含一个Inr,在-137碱基对处界定,在HeLa细胞体内和HeLa细胞提取物体外均具有完全的转录活性。该启动子在淋巴细胞提取物体外也具有高活性,但在不表达内源性TEF-1基因的淋巴细胞系体内则无活性。这个足以在体内指导TEF-1基因组织特异性表达的最小启动子,包含多个结合普遍存在的转录因子Sp1和ATF-1的位点。Inr的突变完全消除了主要起始位点的转录,而次要位点的转录略有增加。近端Sp1位点的失活消除了主要起始位点的转录,并增加了一个5'次要起始位点的转录。插入一个TATA盒元件并没有定性地改变起始位点使用的模式,该模式似乎依赖于上游Sp1位点的完整性。这些观察结果表明Inr和近端元件之间存在“串扰”以确定转录起始位点,这与间距和TATA元件的存在无关。