Structural and functional analysis of hypoxia-inducible factor 1
Structural and functional analysis of hypoxia-inducible factor 1
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DOI：
10.1038/ki.1997.77
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发表时间：
1997-02-01
影响因子：
19.6
通讯作者：
Yu, A
中科院分区：
文献类型：
--
作者：
Semenza, GL;Agani, F;Yu, A
缺氧诱导因子1（HIF-1）是一种碱性螺旋-环-螺旋蛋白，其激活缺氧诱导基因的转录，包括编码以下的基因：促红细胞生成素、血管内皮生长因子、血红素加氧酶-1、诱导型一氧化氮合酶和糖酵解酶醛缩酶A、烯醇化酶1、乳酸脱氢酶A、磷酸果糖激酶L和磷酸甘油酸激酶1。来自这些基因的缺氧反应元件由HIF-1结合位点（其含有核心序列5 '-CGTG-3'）以及功能所需的额外DNA序列组成，所述额外DNA序列在一些元件中包括第二个HIF-1结合位点。HIF-1是一种异源二聚体。HIF-1 α亚基是HIF-1所特有的，而HIF-1 β（ARNT）可以与其他bHLH-PAS蛋白二聚化。HIF-1 α的结构分析显示，与HIF-1 β（ARNT）的二聚化需要HLH和PAS结构域，DNA结合由碱性结构域介导，并且HIF-1 α含有羧基末端反式激活结构域。共转染HIF-1 α和HIF-1 β（ARNT）表达载体和含有野生型缺氧反应元件的报告基因导致非缺氧细胞中转录增加和缺氧细胞中转录的超诱导，而HIF-1表达载体对含有HIF-1结合位点突变的报告基因的转录没有影响。HIF-1 α和HIF-1 β（ARNT）蛋白水平在所有检测的原代和转化细胞系中由缺氧诱导。在HeLa细胞中，HIF-1 α和HIF-1 β蛋白水平以及HIF-1 DNA结合活性随着细胞氧张力降低呈指数增加，最大值在0.5%氧下，半最大值在1.5至2%氧下。HIF-1 α和HIF-1 β（ARNT）mRNA在所有人、小鼠和大鼠器官中检测到，并且在缺氧的啮齿动物中适度诱导mRNA表达。当动物遭受贫血或缺氧时，在体内诱导HIF-1 α蛋白水平。HIF 1A基因定位于人染色体14 q21-q24和小鼠染色体12。
Hypoxia-inducible factor 1 (HIF-1) is a basic helix-loop-helix protein that activates transcription of hypoxia-inducible genes, including those encoding: erythropoietin, vascular endothelial growth factor, heme oxygenase-1, inducible nitric oxide synthase, and the glycolytic enzymes aldolase A, enolase 1, lactate dehydrogenase A, phosphofructokinase L, and phosphoglycerate kinase 1. Hypoxia response elements from these genes consist of a HIF-1 binding site (that contains the core sequence 5'-CGTG-3') as well as additional DNA sequences that are required for function, which in some elements include a second HIF-1 binding site. HIF-1 is a heterodimer. The HIF-1 alpha subunit is unique to HIF-1, whereas HIF-1 beta (ARNT) can dimerize with other bHLH-PAS proteins. Structural analysis of HIF-1 alpha revealed that dimerization with HIF-1 beta (ARNT) requires the HLH and PAS domains, DNA binding is mediated by the basic domain, and that HIF-1 alpha contains a carboxyl-terminal transactivation domain. Co-transfection of HIF-1 alpha and HIF-1 beta (ARNT) expression vectors and a reporter gene containing a wild-type hypoxia response element resulted in increased transcription in non-hypoxic cells and a superinduction of transcription in hypoxic cells, whereas HIF-1 expression vectors had no effect on the transcription of reporter genes containing a mutation in the HIF-1 binding site. HIF-1 alpha and HIF-1 beta (ARNT) protein levels were induced by hypoxia in all primary and transformed cell lines examined. In HeLa cells, the levels of HIF-1 alpha and HIF-1 beta protein and HIF-1 DNA-binding activity increased exponentially as cellular oxygen tension decreased, with maximum values at 0.5% oxygen and half-maximal values at 1.5 to 2% oxygen. HIF-1 alpha and HIF-1 beta (ARNT) mRNAs were detected in all human, mouse, and rat organs assayed and mRNA expression was modestly induced in rodents subjected to hypoxia. HIF-1 alpha protein levels were induced in vivo when animals were subjected to anemia or hypoxia. The HIF1A gene was mapped to human chromosome 14q21-q24 and mouse chromosome 12.