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首页> 美国卫生研究院文献>Turkish Journal of Biology >Identification of IFITM3 and MGAT1 as novel interaction partners of BRI3 by yeast two-hybrid screening
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Identification of IFITM3 and MGAT1 as novel interaction partners of BRI3 by yeast two-hybrid screening

机译：通过酵母双杂交筛选鉴定IFITM3和MGAT1作为BRI3的新型相互作用伴侣

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BRI3 (brain protein I3) is one of the Wnt/β-catenin pathway target genes as indicated by the results of serial analysis of gene expression (SAGE) and microarray analyses performed in our laboratory. The Wnt/β-catenin signaling pathway is an evolutionarily conserved pathway, which has important functions in early vertebrate development, axis formation, cellular proliferation, and morphogenesis. Previous studies showed that BRI3 expression is upregulated at both mRNA and protein levels upon β-catenin activation by various approaches, such as lithium treatment and overexpression of Wnt ligands in Huh7 (hepatocellular carcinoma) cell lines. Moreover, with regard to the previous literature, BRI3 was found to have a very important role in the TNFα-mediated cell death pathway. In this study, we screened a human liver cDNA library by yeast two-hybrid assay using BRI3 protein as bait, with the aim of finding novel interaction partners of BRI3. Library screening by yeast mating resulted in the identification of three candidate positive clones. Among these, IFITM3 and MGAT1 proteins were confirmed as interaction partners by using cotransformation in yeast cells and coimmunoprecipitation from mammalian cell lines. Considering the poor functional characterization of BRI3 to date, identification of novel BRI3-interacting proteins is an essential first step in determining the action mechanism of BRI3 with respect to the Wnt/β-catenin pathway.
机译：BRI3（脑蛋白I3）是Wnt /β-catenin途径的靶基因之一，基因表达的系列分析（SAGE）和我们实验室进行的微阵列分析结果表明。 Wnt /β-catenin信号传导途径是进化保守的途径，在脊椎动物早期发育，轴形成，细胞增殖和形态发生中具有重要功能。先前的研究表明，通过各种方法（例如锂处理和Huh7（肝细胞癌）细胞系中Wnt配体的过表达），β-catenin激活后，BRI3的表达在mRNA和蛋白水平上均被上调。此外，根据先前的文献，发现BRI3在TNFα介导的细胞死亡途径中具有非常重要的作用。在这项研究中，我们以BRI3蛋白为诱饵，通过酵母双杂交法筛选了人肝脏cDNA文库，目的是寻找BRI3的新型相互作用伴侣。通过酵母交配的文库筛选导致鉴定了三个候选阳性克隆。其中，通过使用酵母细胞中的共转化和来自哺乳动物细胞系的共免疫沉淀，证实了IFITM3和MGAT1蛋白是相互作用的伴侣。考虑到迄今为止BRI3的功能特性不佳，鉴定新的BRI3相互作用蛋白是确定BRI3相对于Wnt /β-catenin途径的作用机制的重要第一步。

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