Cloning and functional characterization of the murine caspase-3 gene promoter
Cloning and functional characterization of the murine caspase-3 gene promoter
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DOI：
10.1089/dna.2006.25.104
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发表时间：
2006-02-01
影响因子：
3.1
通讯作者：
Sékaly, RP
中科院分区：
文献类型：
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作者：
Sabbagh, L;Bourbonnière, M;Sékaly, RP
几项研究表明，caspase-3的水平在不同的细胞凋亡条件下上调。以前，我们已经表明，通过TCR激活T细胞导致caspase-3水平上调。这些发现强调了调节caspase-3表达以防止细胞过早死亡的重要性。为了更好地理解caspase-3基因的调控，克隆了5 '-非翻译区的一部分，测序并表征。半胱天冬酶-3基因的5 '侧翼区的片段也被克隆到荧光素酶报告基因的上游，证明该片段含有启动子活性。在Jurkat T细胞中发现了几种启动子缺失构建体的较高荧光素酶表达，但在小鼠Neuro-2A神经母细胞瘤细胞系中没有发现，这表明存在T细胞特异性调节区。这些序列的重要性进一步得到了人类和小鼠胱天蛋白酶-3启动子区域的基因组组织的支持。这些发现表明，caspase-3启动子的-2245/+14区域显示组成型表达水平，并且启动子的几个区域在基础调节中起作用。最后，在人类和小鼠启动子之间鉴定的一些保守的转录因子结合位点似乎在淋巴样细胞中起重要作用。
Several studies have shown that the levels of caspase-3 are upregulated under different conditions of apoptosis. Previously, we have shown that activation of T cells through the TCR leads to the upregulation of caspase-3 levels. These findings highlight the importance of regulating the expression of caspase-3 in order to prevent premature cell death. To better understand the regulation of the caspase-3 gene, a portion of the 5'-untranslated region was cloned, sequenced, and characterized. The segment of the 5'-flanking region of the caspase-3 gene was also cloned upstream of a luciferase reporter gene, demonstrating that this fragment contains promoter activity. Higher luciferase expression was found with several of the promoter deletion constructs in Jurkat T cells but not the mouse Neuro-2A neuroblastoma cell line, suggesting the presence of a T-cell-specific regulated region. The importance of these sequences is further supported by the genomic organization of the human and mouse caspase-3 promoter regions. These findings demonstrated that the -2245/+14 region of the caspase-3 promoter shows constitutive levels of expression, and that several regions of the promoter play a role in basal regulation. Finally, some of the conserved transcription factor binding sites identified between the human and mouse promoters appear to play an important role in lymphoid cells.