Analysis of rainbow trout Ah receptor protein isoforms in cell culture reveals conservation of function in Ah receptor-mediated signal transduction.
Analysis of rainbow trout Ah receptor protein isoforms in cell culture reveals conservation of function in Ah receptor-mediated signal transduction.
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对细胞培养物中虹鳟鱼 Ah 受体蛋白亚型的分析揭示了 Ah 受体介导的信号转导功能的保守性。
DOI：
10.1016/s0006-2952(02)01061-4
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发表时间：
2002
影响因子：
5.8
通讯作者：
Marks-Sojka,Katherine
中科院分区：
文献类型：
--
作者：
Pollenz,RichardS;Necela,Brian;Marks-Sojka,Katherine
从虹鳟鱼中分离出两种不同的芳香烃受体（AHR）cDNA。被称为rtAHR 2 α和rtAHR 2 β的编码受体蛋白产物在氨基酸水平上97%相同，但据报道在AHR介导的基因调控方面具有不同的功能。为了验证这一假设，在体外和中国仓鼠肺细胞系E36中对这两种蛋白质进行了功能评价。为了便于分析，两种rtAHR 2同种型都用FLAG肽标记，并且可以在兔网织红细胞裂解物中表达和定量。然而，这两种蛋白质未能形成功能复合物与哺乳动物或虹鳟鱼AHR核转位蛋白（ARNT），可以与异生素反应元件（XREs）在体外配体依赖性的方式。相反，这两种蛋白质在E36细胞系中表达时，对AHR介导的信号传导表现出正功能。两种rtAHR 2亚型均表现出在未配体状态下的细胞质分布，并且可以在鳟鱼CYP 1A 3启动子的控制下驱动报告基因的表达。虽然两种蛋白质诱导的报告基因活性相同，但两种异构体对2，3，7，8-四氯二苯并-p-二恶英（TCDD）的EC 50值相差一个数量级，rtAHR 2 R异构体对TCDD的反应较低。当rtAHR 2亚型的功能进行了测试的情况下，显性负rtARNTaprotein，TCDD介导的诱导报告基因活性的rtARNTaprotein水平的增加而减少。总之，两种rtAHR 2亚型似乎在AHR介导的信号传导中表现出正功能，表明功能保守。
Two distinct aryl hydrocarbon receptor (AHR) cDNAs have been isolated from rainbow trout. The encoded receptor protein products termed rtAHR2α and rtAHR2ß are 97% identical at the amino acid level but are reported to have distinct functions with regard to AHR-mediated gene regulation. To test this hypothesis, the two proteins were evaluated functionally both in vitro and in a Chinese hamster lung cell line, E36. To facilitate analysis, both rtAHR2 isoforms were tagged with the FLAG peptide and could be expressed and quantified in a rabbit reticulocyte lysate. However, both proteins failed to form functional complexes with mammalian or rainbow trout AHR nuclear translocator protein (ARNT) that could associate with xenobiotic response elements (XREs) in a ligand-dependent manner in vitro. In contrast, both proteins exhibited positive function on AHR-mediated signaling when expressed in the E36 cell line. Both rtAHR2 isoforms showed a cytoplasmic distribution in the unliganded state and could drive the expression of a reporter gene under control of the trout CYP1A3 promoter. Although both proteins induced reporter gene activity to the same magnitude, the EC50values of the two isoforms for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) differed by an order of magnitude, with the rtAHR2ß isoform less responsive to TCDD. When the functions of the rtAHR2 isoforms were tested in the context of the dominant negative rtARNTaprotein, TCDD-mediated induction of reporter gene activity was reduced as the level of rtARNTaprotein increased. In summary, both rtAHR2 isoforms appear to exhibit positive function in AHR-mediated signaling, suggesting conservation of function.
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影响因子：
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作者：
Necela,B;Pollenz,RS
通讯作者：
Pollenz,RS
影响因子：
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作者：
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DOI：
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发表时间：
1997
期刊：
影响因子：
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作者：
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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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影响因子：
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作者：
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通讯作者：
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