Steplewski A, Haas S, Amini S, Khalili K
Jefferson Institute of Molecular Medicine, Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Gene. 1995 Mar 10;154(2):215-8. doi: 10.1016/0378-1119(94)00816-b.
Cell- and stage-specific transcription of the myelin basic protein (MBP)-encoding gene (Mbp) in brain is regulated by arrays of cis-acting regulatory elements positioned at the 5'-flanking region of the gene. The proximal element between nucleotides -14 to -50, termed MB1, has previously been shown to have an important role in the cell-type-specific transcription of Mbp in cells derived from the central nervous system (CNS). Here, we utilized band-shift and in vitro transcription assays to examine the ability of MEF-2, an expression factor encoded by a cDNA isolated from mouse brain, in binding to the MB1 element and regulating transcription of the Mbp promoter. Results from the band-shift assays indicated that the bacterially produced MEF-2 recognizes specific nt in the MB1 motif, and its binding to these nt reduces the overall transcriptional activity of Mbp in a cell-free extract.
大脑中髓鞘碱性蛋白（MBP）编码基因（Mbp）的细胞和阶段特异性转录受位于该基因5'侧翼区域的顺式作用调控元件阵列的调节。核苷酸-14至-50之间的近端元件，称为MB1，先前已被证明在源自中枢神经系统（CNS）的细胞中Mbp的细胞类型特异性转录中起重要作用。在这里，我们利用凝胶迁移和体外转录试验来检测MEF-2（一种从小鼠大脑中分离的cDNA编码的表达因子）与MB1元件结合并调节Mbp启动子转录的能力。凝胶迁移试验结果表明，细菌产生的MEF-2识别MB1基序中的特定核苷酸，并且它与这些核苷酸的结合降低了无细胞提取物中Mbp的整体转录活性。
