Transcriptional regulation of mouse MARCKS promoter in immortalized hippocampal cells.
Transcriptional regulation of mouse MARCKS promoter in immortalized hippocampal cells.
复制标题
永生化海马细胞中小鼠 MARCKS 启动子的转录调控。
DOI：
10.1006/bbrc.2002.6655
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发表时间：
2002
影响因子：
3.1
通讯作者：
Lenox,RobertH
中科院分区：
文献类型：
--
作者：
Wang,Le;Liu,Xingge;Lenox,RobertH
小鼠MARCKS是一种显著的肉豆蔻酰化富含丙氨酸的C激酶底物，与脑发育、钙/钙调蛋白信号传导和膜细胞骨架重构有关，并且以细胞和组织特异性方式发育调节。在这项研究中，小鼠MARCKS启动子在神经源性永生化海马细胞（HN 33）中的转录调控被检测到相对于翻译起始位点的5′侧翼基因组序列的-993到+1部分。在这个神经细胞系中进行的转染实验首次发现，从-993到-713的远端启动子片段在MARCKS核心启动子序列驱动的基础转录活性的上调中作为增强子/激活子元件起着至关重要的作用。基序分析显示，在该区域至少有12个重叠的潜在转录因子结合位点，其中一个突出的富含GA的序列以−765为中心，已被证明在Sp1蛋白样复合物的结合中具有重要的功能。富含GA的片段的缺失显著降低了MARCKS启动子活性。此外，竞争性EMSA表明−993/−713中的两个额外位点也可能与Sp1蛋白相互作用，表明−993/−713的激活功能受多种Sp1转录因子控制。与远端启动子序列不同，近端核心启动子序列（−649/−438）包含一个富含GC的盒和一个Z-DNA形成片段，对基础转录至关重要。缺失−649/−438片段已被证明即使在−993/−713存在的情况下也会严重损害启动子活性，这表明它的存在对−993/−713的功能也很重要。这些数据强调，远端和近端启动子序列之间的协同相互作用是不可或缺的最佳MARCKS启动子在永生化海马细胞的功能。MARCKS远端启动子区激活子功能的发现及其与多种Sp蛋白相互作用的可能性为理解Macs在脑内的转录调控提供了新的线索。
Mouse MARCKS is a prominent myristoylated alanine-rich C kinase substrate implicated in brain development, calcium/calmodulin signaling, and membrane cytoskeletal restructuring, and is developmentally regulated in a cell- and tissue-specific fashion. In this study, transcriptional regulation of mouse MARCKS promoter in the neuronally derived immortalized hippocampal cells (HN33) was examined for a portion of 5′-flanking genomic sequence from −993 to +1 relative to the translation start site. Transfection experiments carried out in this neural cell line identified, for the first time, that the distal promoter segment from −993 to −713 plays a crucial role as an enhancer/activator element in the up-regulation of the basal transcription activity driven by MARCKS core promoter sequence. Motif analyses revealed at least 12 overlapping potential transcription factor binding sites in this region, among which a prominent GA-rich sequence centered at −765 has been shown to be functionally important in the binding of Sp1 protein-like complex. Deletion of the GA-rich segment significantly reduced the MARCKS promoter activity. Further, competitive EMSA indicated two additional sites within the −993/−713 that may also interact with Sp1 protein, demonstrating that the activator function of −993/−713 is under control of multiple Sp1 transcription factors. Unlike the distal promoter sequence, the proximal core promoter sequence (−649/−438) contains a GC-rich box and a Z-DNA-forming segment and is critical to basal transcription. The deletion of −649/−438 segment has been shown to drastically impair the promoter activity even in the presence of −993/−713, suggesting that its presence is also important to the function of −993/−713. These data emphasize that the synergistic interaction between distal and proximal promoter sequences is indispensable for the optimal MARCKS promoter function in the immortalized hippocampal cells. The discovery of the activator function of the MARCKS distal promoter region, and its potential interaction with multiple Sp proteins may provide a new clue to the understanding of Macs transcriptional regulation in brain.
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影响因子：
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作者：
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DOI：
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影响因子：
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作者：
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通讯作者：
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