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当前位置: 首页 > 文章 > 牙鲆甲状腺素受体β互作蛋白的鉴定及其受甲状腺素调控的分析 海洋渔业 2018 (6) 703-712
Position: Home > Articles > Identification of thyroid hormone receptor β-interacting proteins and their regulation by thyroid hormone in Paralichthys olivaceus Marine Fisheries 2018 (6) 703-712

牙鲆甲状腺素受体β互作蛋白的鉴定及其受甲状腺素调控的分析

作  者：
喻杰;付元帅;施志仪
单  位：
上海海洋大学水产与生命学院农业部淡水水产种质资源重点实验室
关键词：
牙鲆;甲状腺素受体β;结合蛋白;免疫沉淀
摘  要：
甲状腺素(TH)在牙鲆(Paralichthys olivaceus)变态发育过程中具有重要的调控作用,其通过与甲状腺素受体结合并在甲状腺素受体结合蛋白的共同作用下发挥生物学功能。利用免疫沉淀和质谱分析法鉴定了牙鲆甲状腺素受体β(TRβ)的结合蛋白,并采用实时荧光定量PCR技术检测了结合蛋白在牙鲆变态过程中受外源甲状腺素调控的变化。从TRβ多克隆抗体与牙鲆胚胎细胞总蛋白的免疫沉淀复合物中鉴定得到了21个牙鲆蛋白,生物信息学分析显示它们参与到多种生命活动中。对其中7个代表性蛋白的基因在出膜后第21天(21 dph)和第28天(28 dph)的牙鲆仔鱼体内的转录水平检测发现,它们的表达在外源甲状腺素处理后均表现出显著变化,其中beta-F1-ATPase、Hsp90β、Cct6A、RPS15A、Slc25a5和EF1a的表达受外源甲状腺素处理显著下调,CK1截然相反,它的表达受甲状腺素处理显著上调;硫脲处理对所分析的基因则几乎都没有明显影响。本研究为进一步探究甲状腺素受体介导甲状腺素调控牙鲆变态提供了新的见解。
译  名：
Identification of thyroid hormone receptor β-interacting proteins and their regulation by thyroid hormone in Paralichthys olivaceus
作  者：
YU Jie;FU Yuan-shuai;SHI Zhi-yi;Key Laboratory of Genetic Resources for Freshwater Aquaculture and Fisheries,College of Fisheries and Life Science,Shanghai Ocen University;
单  位：
YU Jie%FU Yuan-shuai%SHI Zhi-yi%Key Laboratory of Genetic Resources for Freshwater Aquaculture and Fisheries,College of Fisheries and Life Science,Shanghai Ocen University
关键词：
Paralichthys olivaceus;;thyroid hormone receptor β;;interacting proteins;;immunoprecipitation
摘  要：
Thyroid hormone( TH) plays an important role in the metamorphosis of Paralichthys olivaceus. It exerts biological functions through the combination of thyroid hormone receptors together with thyroid hormone receptor interacting proteins. In this study,immunoprecipitation and mass spectrometry were used to identify the interacting proteins of Paralichthys olivaceus thyroid hormone receptor β,and real-time fluorescence quantitative PCR was used to detect the changes of the interacting proteins in the metamorphosis of Paralichthys olivaceus affected by exogenous thyroid hormone at 21 dph and 28 dph. We identified 21 Paralichthys olivaceus proteins from immunoprecipitated complexes of TRβ polyclonal antibodies and total Paralichthys olivaceus embryonic cell proteins. Bioinformatics analysis revealed that they participated in various life activities. The mRNA expression levels of seven representative proteins were detected and their expressions showed significant changes after exogenous thyroid hormone treatment. The expression of beta-F1-ATPase, Hsp90β, Cct6 A, RPS15 A, Slc25 a5, and EF1 a was significantly downregulated by exogenous thyroid hormone treatment. In contrast,CK1 was significantly up-regulated by thyroid hormone treatment,and thiourea treatment had almost no significant effect on the analyzed genes. This study provides new insights into the role of thyroid hormone receptor-mediated regulation of thyroid hormone in Paralichthys olivaceus metamorphosis.

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