Asaka Masamitsu N, Murano Kensaku, Nagata Kyosuke
Department of Infection Biology, Graduate School of Comprehensive Human Science and Institute of Basic Medical Science, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan.
Biochem Biophys Res Commun. 2008 Nov 28;376(4):665-70. doi: 10.1016/j.bbrc.2008.09.053. Epub 2008 Sep 20.
TAF-I, one of histone chaperones, consists of two subtypes, TAF-Ialpha and TAF-Ibeta. The histone chaperone activity of TAF-I is regulated by dimer patterns of these subtypes. TAF-Ibeta is expressed ubiquitously, while the expression level of TAF-Ialpha with less activity than TAF-Ibeta differs among cell types. It is, therefore, assumed that the expression level of TAF-Ialpha in a cell is important for the TAF-I activity level. Here, we found that TAF-Ialpha and TAF-Ibeta genes are under the control of distinct promoters. Reporter assays and gel shift assays demonstrated that Sp1 binds to three regions in the TAF-Ialpha promoter and two or all mutaions of the three Sp1 binding regions reduced the TAF-Ialpha promoter activity. ChIP assays demonstrated that Sp1 binds to the TAF-Ialpha promoter in vivo. Furthermore, the expression level of TAF-Ialpha mRNA was reduced by knockdown of Sp1 using siRNA method. These studies indicated that the TAF-Ialpha promoter is under the control of Sp1.
TAF-I是组蛋白伴侣之一,由两种亚型TAF-Iα和TAF-Iβ组成。TAF-I的组蛋白伴侣活性受这些亚型二聚体模式的调节。TAF-Iβ广泛表达,而活性低于TAF-Iβ的TAF-Iα的表达水平在不同细胞类型中有所不同。因此,推测细胞中TAF-Iα的表达水平对TAF-I活性水平很重要。在此,我们发现TAF-Iα和TAF-Iβ基因受不同启动子的控制。报告基因分析和凝胶迁移分析表明,Sp1与TAF-Iα启动子中的三个区域结合,这三个Sp1结合区域的两个或全部突变会降低TAF-Iα启动子活性。染色质免疫沉淀分析表明,Sp1在体内与TAF-Iα启动子结合。此外,使用小干扰RNA方法敲低Sp1可降低TAF-Iα mRNA的表达水平。这些研究表明,TAF-Iα启动子受Sp1的控制。