Identification and characterization of the interaction between tuberin and 14-3-3ζ
Identification and characterization of the interaction between tuberin and 14-3-3ζ
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DOI：
10.1074/jbc.m204802200
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发表时间：
2002-10-18
影响因子：
4.8
通讯作者：
Halley, DJJ
中科院分区：
文献类型：
--
作者：
Nellist, M;Goedbloed, MA;Halley, DJJ
脑硬化症是由肿瘤抑制基因TSC 1或TSC 2突变引起的。该疾病的特征在于广泛的表型谱，包括癫痫发作、精神发育迟滞、肾功能障碍和皮肤病学异常。TSC 1编码一种130 kDa的蛋白质，称为hamartin，TSC 2编码一种200 kDa的蛋白质，称为tuberin。虽然已经表明错构蛋白和块茎蛋白形成复合物并介导核糖体蛋白S6的磷酸肌醇3-激酶/Akt依赖性磷酸化，但尚不清楚任一蛋白的失活如何导致结节性硬化症。因此，为了获得对块茎蛋白和错构蛋白功能的额外了解，进行酵母双杂交筛选实验以鉴定与块茎蛋白相互作用的蛋白质。其中一种蛋白质是14-3- 3 zeta，是14-3- 3 zeta蛋白家族的成员。在体外和通过免疫共沉淀证实了结核菌素和14-3- 3 zeta之间的相互作用;鉴定了结核菌素内用于14-3- 3 zeta结合的多个位点;并且确定14-3- 3 zeta与结核菌素-错构瘤蛋白复合物相关。最后，显示了块茎蛋白/14-3- 3 zeta相互作用由Akt介导的块茎蛋白磷酸化调节，提供了对块茎蛋白如何调节S6磷酸化的深入了解。
Tuberous sclerosis is caused by mutations to either the TSC1 or TSC2 tumor suppressor gene. The disease is characterized by a broad phenotypic spectrum that includes seizures, mental retardation, renal dysfunction, and dermatological abnormalities. TSC1 encodes a 130-kDa protein called hamartin, and TSC2 encodes a 200-kDa protein called tuberin. Although it has been shown that hamartin and tuberin form a complex and mediate phosphoinositide 3-kinase/Akt-dependent phosphorylation of the ribosomal protein S6, it is not yet clear how inactivation of either protein leads to tuberous sclerosis. Therefore, to obtain additional insight into tuberin and hamartin function, yeast two-hybrid screening experiments were performed to identify proteins that interact with tuberin. One of the proteins identified was 14-3-3zeta, a member of the 14-3-3zeta protein family. The interaction between tuberin and 14-3-3zeta was confirmed in vitro and by co-immunoprecipitation; multiple sites within tuberin for 14-3-3zeta binding were identified; and it was determined that 14-3-3zeta associated with the tuberin-hamartin complex. Finally, it was shown that the tuberin/14-3-3zeta interaction is regulated by Akt-mediated phosphorylation of tuberin, providing insight into how tuberin may regulate phosphorylation of S6.