Gene Expression of Cell-adhesive Molecule Vitronectin

1.小鼠黑色素瘤细胞整合素α v顺式作用元件的鉴定。(Head我们从小鼠基因组文库中克隆了小鼠整联蛋白启动子区，并确定了转录起始位点上游1.8 kbp区域（+1 bp）的核苷酸序列。我们已经揭示了-108和+1 bp之间的三个转录调控元件。其中一个片段（-27 ~-16 bp）与Sp1结合，另外两个片段（-61 ~-54 bp和-106 ~-98 bp）与Ets家族结合。从-61到-54 bp的元件与Ets家族的Ets-1结合。3个顺式作用元件中任何一个的突变或缺失都显著降低了启动子活性。这些结果表明Sp1和Ets协同激活109 bp的α v启动子区域.玻连蛋白启动子区域的表征（研究者，Yasunori Miyamoto博士）我们表明，-19至+54 bp的区域对于小鼠玻连蛋白基因中的转录起始是必需的。此区域没有TATA框。它与从人肝癌HepG 2细胞中提取的核蛋白形成复合物。我们测定了玻连蛋白基因5 ′侧翼区-1.8 ~-3.7kbp的核苷酸序列。

1. Identification of cis-acting elements of integrin α v in mouse melanoma cells. (Head Investigator, Prof. Masao Hayashi)We have cloned mouse integrin promoter region from mouse genome library and determined the nucleotide sequence of 1.8 kbp region upstream of the transcription initiation site (+1 bp). We have revealed the three transcriptional regulatory elements between -108 and +1 bp. One element from -27 to -16 bp binds to Sp1, and the two other elements from -61 to -54 bp and from -106 to -98 bp bind to Ets family. The element from -61 to -54 bp binds to Ets-1, one of the Ets family. Mutations or deletions in any one of the 3 cis-acting elements remarkably decreased the promoter activity. These results indicate that Sp1 and Ets cooperate to activate the 109-bp α v promoter region.2. Characterization of vitronectin promoter region (Investigator, Dr. Yasunori Miyamoto)We showed that a region from -19 to +54 bp is essential for the initiation of transcription in mouse vitronectin gene. This region does not have TATA box. It forms a complex with nuclear proteins extracted from human hepatoma HepG2 cells. We determined the nucleotide sequence between -1.8 kbp and -3.7 kbp in 5'-flanking region of vitronectin gene.