【摘要】 目的寻找与近日节律系统核心基因Period1(Per1)相互作用的新蛋白,揭示近日节律相关的可能信号通路。方法采用酵母双杂交方法,以人Per1的PAS结构域为诱饵,扫描人下丘脑cDNA文库,并通过体外转录、免疫共沉淀验证蛋白间相互作用;通过RT-PCR检测RACK1(receptors for activated C-k inase)表达的组织特异性及节律性;运用RNA i技术初步研究RACK1与Per1的信号联系。结果人下丘脑区域RACK1蛋白与Per1存在相互作用;RACK1在多器官组织保守表达,但其表达未呈现明显节律性;上调及下调Per1表达,RACK1的RNA水平无显著变化。结论细胞内接头分子RACK1是一种新的Per1作用蛋白,可能介导、调控Per1的功能。更多还原

【Abstract】 Objective To identify novel proteins interacting with Period1（Per1）,which is a core component of the circadian clock system,and to uncover intracellular signal pathway devoted to circadian clock. Method Using hPer1_PAS domain as bait,human brain cDNA library was scanned by yeast two-hybrid.The interaction was confirmed by transcription/translation in vitro and co-immunoprecipitation tests.RT-PCR was used to investigate the expressional characteristic of RACK1（receptors for activated C-kinase） including tissue specificity and rhythmicity.The signal relation between RACK1 and Per1 was studied by RNA-interference.Result Per1 interplayed with RACK1 in the human SCN region of the hypothalamus.The expression of RACK1 was conserved in various human organs and did not display a robust and stable pattern of rhythmicity in mouse brain. Up-and down-regulating of Per1 had no significant effects on the RACK1 expression in cell tests.Conclusion The special interaction between RACK1 and Per1 has been approved,which means RACK1 may serve as an adaptor molecule to modulate function of hPer1.更多还原