HUMAN TENASCIN GENE - STRUCTURE OF THE 5'-REGION, IDENTIFICATION, AND CHARACTERIZATION OF THE TRANSCRIPTION REGULATORY SEQUENCES
HUMAN TENASCIN GENE - STRUCTURE OF THE 5'-REGION, IDENTIFICATION, AND CHARACTERIZATION OF THE TRANSCRIPTION REGULATORY SEQUENCES
复制标题
DOI：
10.1074/jbc.270.7.3429
复制
发表时间：
1995-02-17
影响因子：
4.8
通讯作者：
ZARDI, L
中科院分区：
文献类型：
--
作者：
GHERZI, R;CARNEMOLLA, B;ZARDI, L
本文描述了人TN基因5‘端区域的基因组结构及其启动子的功能特征。TN基因5‘侧翼区约2300个碱基对已被克隆和测序。这个基因组区域包含几个潜在的转录因子结合位点。通过引物延伸和S1核酸酶分析，我们定位了转录起始点。TN基因的第一个外显子(179个碱基对长)存在于两个主要的TN转录本中，表明这两个mRNAs的表达受单个启动子的调控。转录起始点上游的220个碱基在TN产生细胞和非产生细胞中同样有效地指导氯霉素乙酰转移酶(CAT)报告基因的表达。利用人类5‘-侧翼区的缺失片段，我们已经证明在TN产生和非产生细胞系中都存在-220到-2300区域活跃的假定的’‘沉默’‘元件。此外，我们已经证明，在TN产生细胞中的选择性转录需要在CAT表达载体中存在TN基因内含子1的1.3kb部分。这些发现表明，复杂的机制控制着TN基因的转录调控。
This report describes the genomic organization of the 5'-region of the human tenascin-C (TN) gene and the functional characterization of its promoter. Approximately 2300 base pairs of the TN gene 5'-flanking region have been cloned and sequenced. This genomic region contains several potential binding sites for transcription factors. By primer extension and S1 nuclease analysis we have localized the transcription start site. The first exon of the TN gene (179 base pairs long) is present in the two major TN transcripts, showing that the expression of these two mRNAs is regulated by a single promoter. The 220 bases upstream to the transcription start site are equally active in directing the expression of chloramphenicol acetyltransferase (CAT) reporter gene in TN producer and nonproducer cells. Using deletion fragments of the human 5'-flanking region we have shown the presence of putative ''silencer'' elements in the -220 to -2300 region active in both TN producer and nonproducer cell lines. Furthermore, we have demonstrated that the selective transcription in TN producing cells requires the presence of a 1.3-kilobase portion of the TN gene intron 1 in the CAT expression vectors. These findings indicate that complex mechanisms control the transcriptional regulation of TN gene.