A cis-acting element in the promoter region of the murine c-myc gene is necessary for transcriptional block.
A cis-acting element in the promoter region of the murine c-myc gene is necessary for transcriptional block.
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鼠 c-myc 基因启动子区域的顺式作用元件对于转录阻断是必需的。
DOI：
10.1128/mcb.9.12.5340-5349.1989
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发表时间：
1989
影响因子：
5.3
通讯作者：
Nepveu,A
中科院分区：
文献类型：
--
作者：
Miller,H;Asselin,C;Dufort,D;Yang,JQ;Gupta,K;Marcu,KB;Nepveu,A
已证明在人和鼠的mycgenes的第一个外显子内发生转录延长的阻断。发现这种阻断的程度随细胞的生理状态而变化，表明转录阻断的调节可以用于控制该基因的表达。为了确定转录阻断所需的切割序列，我们构建了一系列含有murinec-myc 5 ′-侧翼和外显子1序列的构建体。我们建立了用这些构建体稳定转染的HeLa和CV-1细胞群体。通过SI核酸酶图谱分析确定转录起始位点，并通过核连续转录测定测量转录阻断的程度。我们的研究结果表明，至少有两个顺式作用元件是必要的转录块。一个3′元件被发现位于转录停止的区域，并显示出一些原核生物中发现的终止位点的特征。在P1和P2转录起始位点之间有一个5′元件。我们最近发现，两个核因子结合位点位于Pt和P2之间（C。Asselin，A. Nepveu和K. B。Marcu，Oncogene 4：549-558，1989）。去除更多的3′结合位点消除了转录阻断。
A block to elongation of transcription has been shown to occur within the first exon of the human and murinec-mycgenes. The extent of this block was found to vary with the physiological state of cells, indicating that modulation of the transcriptional block can serve to control the expression of this gene. To determine which sequences are required incisfor the transcriptional block, we generated a series of constructs containing various portions of murinec-myc5′-flanking and exon 1 sequences. We established populations of HeLa and CV-1 cells stably transfected with these constructs. The transcription start sites were determined by SI nuclease mapping analysis, and the extent of transcriptional block was measured by nuclear run-on transcription assays. Our results demonstrate that at least two cw-acting elements are necessary for the transcriptional block. A 3′ element was found to be located in the region where transcription stopped and showed features reminiscent of some termination sites found in procaryotes. A 5′ element was positioned between the P1and P2transcription start sites. We recently demonstrated that two nuclear factor-binding sites reside between Ptand P2(C. Asselin, A. Nepveu, and K. B. Marcu, Oncogene 4:549-558, 1989). Removal of the more 3′ binding site abolished the transcriptional block.