Hateboer G, Timmers H T, Rustgi A K, Billaud M, van 't Veer L J, Bernards R
Division of Molecular Genetics, Massachusetts General Hospital, Charlestown.
Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8489-93. doi: 10.1073/pnas.90.18.8489.
Using a protein binding assay, we show that the amino-terminal 204 amino acids of the c-Myc protein interact directly with a key component of the basal transcription factor TFIID, the TATA box-binding protein (TBP). Essentially the same region of the c-Myc protein also binds the product of the retinoblastoma gene, the RB protein. c-Myc protein coimmunoprecipitates with TBP in lysates of mammalian cells, demonstrating that the proteins are also complexed in vivo. A short peptide that spans the RB binding site of the E7 protein of human papilloma virus type 16 interferes with the binding of c-Myc to TBP. The same peptide also blocks binding of adenovirus E1A protein to TBP, suggesting that c-Myc and E1A bind to RB and TBP through overlapping epitopes. Furthermore, we show that binding of RB to E1A prevents association of E1A with TBP. Our data suggest that one of the functions of RB and RB-like proteins is to prevent interaction of viral and cellular oncoproteins, such as c-Myc and E1A, with TBP.
通过蛋白质结合分析,我们发现c-Myc蛋白的氨基末端204个氨基酸直接与基础转录因子TFIID的关键组分——TATA盒结合蛋白(TBP)相互作用。c-Myc蛋白的基本相同区域也与视网膜母细胞瘤基因的产物RB蛋白结合。在哺乳动物细胞裂解物中,c-Myc蛋白与TBP共免疫沉淀,表明这些蛋白在体内也形成复合物。一段跨越人乳头瘤病毒16型E7蛋白RB结合位点的短肽可干扰c-Myc与TBP的结合。同一肽段也可阻断腺病毒E1A蛋白与TBP的结合,这表明c-Myc和E1A通过重叠表位与RB和TBP结合。此外,我们发现RB与E1A的结合可阻止E1A与TBP的结合。我们的数据表明,RB和RB样蛋白的功能之一是阻止病毒和细胞癌蛋白(如c-Myc和E1A)与TBP相互作用。