Nelbock P, Dillon P J, Perkins A, Rosen C A
Department of Molecular Oncology and Virology, Roche Institute of Molecular Biology, Hoffmann-La Roche Inc., Nutley, NJ 07110.
Science. 1990 Jun 29;248(4963):1650-3. doi: 10.1126/science.2194290.
The human immunodeficiency virus (HIV) tat protein (Tat) is a positive regulator of virus gene expression and replication. Biotinylated Tat was used as a probe to screen a lambda gt11 fusion protein library, and a complementary DNA encoding a protein that interacts with Tat was cloned. Expression of this protein, designated TBP-1 (for Tat binding protein-1), was observed in a variety of cell lines, with expression being highest in human cells. TBP-1 was localized predominantly in the nucleus, which is consistent with the nuclear localization of Tat. In cotransfection experiments, expression of TBP-1 was able to specifically suppress Tat-mediated transactivation. The strategy described may be useful for direct identification and cloning of genes encoding proteins that associate with other proteins to modulate their activity in a positive or negative fashion.
人类免疫缺陷病毒（HIV）反式激活因子蛋白（Tat）是病毒基因表达和复制的正向调节因子。生物素化的Tat被用作探针来筛选λgt11融合蛋白文库，并克隆了一个编码与Tat相互作用的蛋白质的互补DNA。这种蛋白质命名为TBP-1（Tat结合蛋白-1），在多种细胞系中均有表达，在人类细胞中表达量最高。TBP-1主要定位于细胞核，这与Tat的核定位一致。在共转染实验中，TBP-1的表达能够特异性抑制Tat介导的反式激活。所述策略可能有助于直接鉴定和克隆编码与其他蛋白质以正向或负向方式结合来调节其活性的蛋白质的基因。
