Two activator protein-1 elements in the matrix metalloproteinase-1 promoter have different effects on transcription and bind Jun D, c-Fos, and Fra-2.
Two activator protein-1 elements in the matrix metalloproteinase-1 promoter have different effects on transcription and bind Jun D, c-Fos, and Fra-2.
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基质金属蛋白酶 1 启动子中的两个激活蛋白 1 元件对转录有不同的影响，并结合 Jun D、c-Fos 和 Fra-2。
DOI：
10.1016/s0945-053x(05)80014-9
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Brinckerhoff,CE
中科院分区：
文献类型：
--
作者：
White,LA;Brinckerhoff,CE
胶原酶，即基质金属蛋白酶-1，作为唯一能在中性pH条件下降解间质胶原蛋白的酶，在结缔组织代谢中起着核心作用。我们使用兔胶原酶启动子的1800到182bp的片段来测量−77处激活蛋白-1(AP-1)位点的转录活性。该序列中−77位的突变极大地降低了所有构建物中的基础转录。然而，启动子至少有321个碱基的突变结构对佛波醇肉豆蔻酸醋酸酯的反应，与它们的天然对应结构相似，暗示上游区域参与了这一反应。通过突变和DNA-蛋白质相互作用的分析，我们还在−第186位鉴定和鉴定了一个新的AP-1位点。启动子321bp−第186位突变略微降低了基本活性，但与−77位突变相比，佛波尔反应性降低了50%。迁移率改变分析显示这两个位点都有特异的可诱导结合。两个AP-1位点的DNA/蛋白质复合体都含有c-Fos和Jun D蛋白，而FrA-2仅存在于−77位点。这些研究(1)证实了这两个AP-1位点之间的协同作用，(2)证实了−186位点在佛波醇诱导中的作用，(3)确定了与这些位点结合的Fos和Jun家族的特定成员。
Collagenase (matrix metalloproteinase-1, MMP-1) plays a central role in connective tissue metabolism as the only enzyme capable of degrading interstitial collagens at neutral pH. We used fragments of the rabbit collagenase promoter ranging from 1800 to 182 bp to measure transcriptional activity of the activator protein-1 (AP-1) site at −77. Mutation at −77 in this sequence greatly reduced basal transcription in all constructs. However, mutant constructs with at least 321 bp of promoter responded to phorbol myristate acetate, similar to their native counterparts, implicating upstream regions in mediating this response. Through mutagenesis and analysis of DNA-protein interactions, we also identified and characterized a novel AP-1 site at −186. Mutation at −186 in 321 bp of promoter modestly lowered basal activity but, in contrast to mutation at −77, reduced phorbol responsiveness by 50%. Mobility shift assays demonstrated specific inducible binding at both sites. DNA/protein complexes at both AP-1 sites contain c-Fos and Jun D proteins, while Fra-2 is present only at the −77 site. These studies (1) demonstrate cooperativity between these two AP-1 sites, (2) implicate the −186 site in phorbol inducibility and (3) identify specific members of the Fos and Jun families binding to these sites.