原癌基因LMO3相互作用蛋白的筛选及鉴定

目的 通过筛选LMO3的相互作用蛋白，进一步了解LMO3的作用及可能机制。方法酵母双杂交方法筛选LMO3相瓦作用蛋白，并通过酵母结合试验、免疫共沉淀及荧光共定位等进行验证。结果在初步获得相互作用蛋白：钙-整合素结合蛋白（Calcium—and integrin—binding protein，CIB）的基础上，在酵母中证实了LMO3与CIB的相互作用，并通过酵母结合试验确定了CIB与LMO3的相互作用位点，发现CIB可与LMO3的第一个LIM结构域（LIMI）及全长LMO3结合，免疫共沉淀试验确证了它们可以在真核细胞内结合，荧光共定位表明与CIB的相互作用可使LMO3在C8细胞中的定位由细胞核移到细胞质。结论LMO3可以与CIB在真核细胞中发生相互作用，提示LMO3可能通过与CIB的相互作用参与细胞相关功能的调节。

Screen and Identification of Proteins Interacting with Proto-oncogene LMO3

Objective This study was aimed to screen LMO3 interacting proteins and get new insight into mechanisms of LMO3. Methods The yeast two-hybrid system was used to isolate and identify proteins that interact with LMO3. The interaction between LMO3 and CIB （calcium-and inte- grin-binding protein） was further confirmed by yeast mating, immunoprecipitation and co-localization assay. Results The CIB was identified as an LMO3 interaction protein. Yeast mating proved that LMO3 interacts with CIB through LIM1 of LMO3 but not LIM2 domain. The interaction of LMO3 and CIB was further confirmed by immunopreeipitation and co-localization assay. LMO3 is co-localized with CIB in the cytoplasm of C8 cells, which results in nuclear transloeation of LMO3. Conclusion LMO3 may interact with CIB in eukaryocytes, suggesting that LMO3 may function through interaction with CIB.