Cloning and functional analysis of the mouse 5-lipoxygenase promoter.
Cloning and functional analysis of the mouse 5-lipoxygenase promoter.
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小鼠5-脂氧合酶启动子的克隆和功能分析。
DOI：
10.1165/ajrcmb.26.4.4747
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发表时间：
2002
影响因子：
6.4
通讯作者：
Lu,Weining
中科院分区：
文献类型：
--
作者：
Silverman,EricS;Le,Louis;Baron,RebeccaM;Hallock,Arlene;Hjoberg,Josephine;Shikanai,Toshiki;Stormvan'sGravesande,Karin;Auron,PhilipE;Lu,Weining
5-脂氧合酶（ALOX 5）是一种对所有白三烯的形成至关重要的酶，在多个水平上受到高度调节，包括基因转录。人ALOX 5启动子序列已被克隆并得到了很好的表征。已鉴定出几个重要的顺式作用元件，包括位于ATG起始密码子上游约145-179个碱基对（bp）的富含G + C的序列。该区域包含转录因子血清蛋白1、锌指转录因子（SP1）和早期生长反应蛋白1、锌指转录因子（EGR-1）的共识结合位点，并且在功能上显著的多态性改变这些序列方面是独特的。为了进一步理解这些多态性和启动子中的其它调控序列的意义，我们从129/SvJ BAC文库中克隆了约2,000 bp的小鼠启动子序列，用于与人基因直接比较。与人类启动子一样，小鼠Alox 5启动子缺乏TATA盒，并具有多个起始位点。紧邻翻译起始位点上游的前292 bp起核心启动子的作用，其能够介导RAW细胞但不是3 T3细胞中的高基础转录。人类和小鼠基因之间的共有元件分布存在巨大差异;然而，存在三个强同源性区域，并且它们包含转录因子的SP1、加塔、GGAGA和ETS家族的共有结合位点。 我们发现，Sp1/Sp3是必不可少的组成型启动子-报告活性。
5-Lipoxygenase (ALOX5), an enzyme essential for the formation of all leukotrienes, is highly regulated at multiple levels, including gene transcription. The humanALOX5promoter sequence has been cloned and is well characterized. Several importantcis-acting elements have been identified including a G + C-rich sequence ∼ 145–179 base pairs (bp) upstream from the ATG start codon. This region contains consensus-binding sites for the transcription factor serum protein 1, a zinc-finger transcription factor (SP1) and early growth-response protein 1, a zinc-finger transcription factor (EGR-1) and is unique in that functionally significant polymorphisms alter these sequences. To further understand the significance of these polymorphisms and other regulatory sequences in the promoter we cloned ∼ 2,000 bp of the mouse promoter sequence from a 129/SvJ BAC library for direct comparison with the human gene. Like the human promoter, the mouse Alox5 promoter lacks a TATA box and has multiple start sites. The first 292 bp immediately upstream of the translational start site function as a core promoter that is capable of mediating high basal transcription in RAW cells but not 3T3 cells. There are vast differences in the distribution of consensusciselements between human and mouse genes; however, three areas of strong homology exist and they contain consensus-binding sites for the SP1, GATA, GGAGA, and ETS family of transcription factors. We show that Sp1/Sp3 is essential for constitutive promoter-reporter activity.