Induction of Human Prointerleukin 1beta Gene by Human T-Cell Leukemia Virus

人T细胞白血病病毒诱导人白介素原1β基因

基本信息

批准号：
08671265
负责人：
TSUKADA Junichi
金额：
$ 1.41万
依托单位：
University of Occupational and Environmental Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671265/
关键词：
Tax HTLV-I transcription Interleukin 1 インターロイキン 1

项目摘要

The human T-cell leukemia virus type I (HTLV-I) , which infects a wide variety of mammalian cells including monocytes and macrophages, encodes a transactivating protein designated as Tax. We now report that Tax induces the human proIL-1beta (IL1B) gene promoter in monocytic cells. In our transient transfection assays using human THP-1 monocytic cells, a chloramphenicol acetyltransferase (CAT) construct containing the IL1B promoter sequence between positions-131 and+12 showed an approximately 90-fold increase in activity following cotransfection of a Tax expression vector. Moreover, Tax synergized with LPS to induce the IL1B promoter activity. Analyzes of specific nucleotide substitutions further indicated that the Tax-induced transcriptional activation requires two transcription factor binding motifs within the IL1B promoter ; One is a binding site for NF-IL6 (CCAAT/enhancer binding protein beta C/EBPbeta) , which belongs to the basic region-leucine zipper (bZIP) family and the other f … More or Spi-1 (PU.1) , which is an Ets family protein found principally in monocytes, macrophages and B lymphocytes. In electrophoretic mobility shift assays (EMSA) using in vivo THP-1 nuclear extracts, Tax expression in THP-1 monocytic cells significantly increased binding of the two factors to their target IL1B promoter sequences. However, in contrast to NF-IL6 and Spi-1, DNA binding activity of Oct-1, a ubiquitously expressed octamer-binding protein was not affected by Tax. Additional EMSA using in vitro translated proteins also showed that recombinant Tax enhances DNA binding of both of recombinant NF-IL6 and Spi-1 proteins. These data were supported by our glutathione S-transferase (GST) -pulldown data, which indicated that Tax physically interacts with the two proteins. Based upon the results obtained from the present study, we conclude that the IL1B promoter is a Tax-responsive sequence as a result of ability of Tax to induce binding of NF-IL6 and Spi-1 to IL1B promoter sequence through protein-protein interaction. Less

人类T细胞白血病病毒I型（HTLV-I）感染了包括单核细胞和巨噬细胞在内的各种哺乳动物细胞，编码了指定为税收的反式激活蛋白质。现在，我们报告说，税收诱导人类proil-1beta（IL1B）基因启动子在单核细胞中。在我们使用人THP-1单核细胞的瞬时转染测定中，氯霉素乙酰基转移酶（CAT）构建体含有位置131和+12之间的IL1B启动子序列，显示出税务表达载体后的活性增加了约90倍。此外，税收与LP协同诱导IL1B启动子活性。对特定核苷酸取代的分析进一步表明，税收诱导的转录激活需要IL1B启动子内的两个转录因子结合基序。一个是NF-IL6（CCAAT/增强剂结合蛋白βC/EBPBETA）的结合位点，属于基本的区域 - 亮氨酸拉链（BZIP）家族和其他F…More或Spi-1（PU.1），这是Monocytes，Mocrophages和Blymphopys and Blymphopytes and Blymphocytes and spi-1（pu.1）。在使用体内THP-1核提取物的电泳迁移率转移测定法（EMSA）中，THP-1单核细胞中的税收表达显着增加了这两个因素与目标IL1B启动子序列的结合。然而，与NF-IL6和SPI-1相比，OCT-1的DNA结合活性（普遍表达的八聚体结合蛋白）不受税收影响。使用体外翻译蛋白的其他EMSA还表明，重组税增强了重组NF-IL6和SPI-1蛋白的DNA结合。这些数据得到了我们的谷胱甘肽S-转移酶（GST）-Pulldown数据的支持，这表明税收与两种蛋白质物理相互作用。根据从本研究中获得的结果，我们包括IL1B启动子是税收响应序列，这是由于税收通过蛋白质 - 蛋白质相互作用诱导NF-IL6和SPI-1与IL1B启动子序列的结合的能力。较少的