Identification of interacting proteins with aryl hydrocarbon receptor in scallop Chlamys farreri by yeast two hybrid screening
Identification of interacting proteins with aryl hydrocarbon receptor in scallop Chlamys farreri by yeast two hybrid screening
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DOI：
10.1016/j.ecoenv.2016.07.013
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发表时间：
2016-11-01
影响因子：
6.8
通讯作者：
Liu, Tong
中科院分区：
文献类型：
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作者：
Cai, Yuefeng;Pan, Luqing;Liu, Tong
芳烃受体（AhR）属于转录因子的基本螺旋环螺旋（bHLH）Per-Arnt-Sim（PAS）家族。AhR主要因其在几种药物和异生物质代谢酶的调节中的作用以及某些异生物质毒性的介导而闻名，包括2，3，7，8-四氯二苯并-对-二恶英（TCDD）。虽然AhR作为海洋双壳贝类中某些外源性物质毒性的介体被充分研究，但其正常的生理功能仍然未知。为了研究AhR的功能，构建了栉孔扇贝（Chlamys farreri）鳃组织cDNA文库和诱饵蛋白表达质粒pGBKT 7-CfAhR。利用酵母双杂交系统，经过高筛选率培养基的多重筛选、旋转验证、测序和生物信息学分析，研究了CfAhR与活化蛋白激酶C1受体（RACK 1）、甲状腺过氧化物酶样蛋白（TPO）、Toll样受体4（TLR 4）、类雄球蛋白、钙库操纵的钙内流（SocE）、ADP/ATP载体蛋白、细胞色素B、硫酯酶、肌动蛋白、铁蛋白亚基1、多聚泛素、短链胶原C4样蛋白和一种假设蛋白在鳃细胞中被鉴定。本研究表明CfAhR在免疫系统稳态、氧化应激反应和栉孔扇贝生长发育中起重要作用。这些蛋白质相互作用的阐明是非常重要的，无论是在理解正常的生理功能的AhR，并作为潜在的目标，进一步研究蛋白质功能的AhR相互作用。(C)2016 Elsevier Inc. All rights reserved.
The aryl hydrocarbon receptor (AhR) belongs to the basic-helix-loop helix (bHLH) Per-Arnt-Sim (PAS) family of transcription factors. AhR has been known primarily for its role in the regulation of several drug and xenobiotic metabolizing enzymes, as well as the mediation of the toxicity of certain xenobiotics, including 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Although the AhR is well-studied as a mediator of the toxicity of certain xenobiotics in marine bivalves, the normal physiological function remains unknown. In order to explore the function of the AhR, the bait protein expression plasmid pGBKT7-CfAhR and the cDNA library of gill from Chlamys farreri were constructed. By yeast two hybrid system, after multiple screening with the high screening rate medium, rotary verification, sequencing and bioinformatics analysis, the interactions of the CfAhR with receptor for activated protein kinase C 1 (RACK1), thyroid peroxidase-like protein (TPO), Toll-like receptor 4(TLR 4), androglobin-like, store-operated Ca2+ entry (SocE), ADP/ATP carrier protein, cytochrome b, thioesterase, actin, ferritin subunit 1, poly-ubiquitin, short-chain collagen C4-like and one hypothetical protein in gill cells were identified. This study suggests that the CfAhR played fundamental roles in immune system homeostasis, oxidative stress response, and in grow and development of C farreri. The elucidation of these protein interactions is of much importance both in understanding the normal physiological function of AhR, and as potential targets for further research on protein function in AhR interactions. (C) 2016 Elsevier Inc. All rights reserved.