Fu G K, Grosveld G, Markovitz D M
Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor 48109-0642, USA.
Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):1811-5. doi: 10.1073/pnas.94.5.1811.
The product of the dek oncogene is the 43-kDa DEK nuclear protein. DEK was first identified in a fusion with the CAN nucleoporin protein in a specific subtype of acute myelogenous leukemia. DEK has also been shown to be an autoantigen in patients with pauciarticular onset juvenile rheumatoid arthritis. Further, the last 65 amino acids of DEK can partially reverse the mutation-prone phenotype of cells from patients with ataxia-telangiectasia. However, in spite of these significant disease associations, the function of DEK has remained unclear. The HIV-2 peri-ets (pets) site is a TG-rich element found between the two Elf-1 binding sites in the HIV-2 enhancer. The pets element mediates transcriptional activation whether the enhancer is stimulated by phorbol 12-myristate 13-acetate (PMA) alone, phytohemagluttinin (PHA) alone, PMA plus PHA, soluble antibodies to the T cell receptor, immobilized antibodies to the T cell receptor, or by antigen. Previously, we purified and characterized the pets factor, demonstrating that it is a 43-kDa nuclear protein. We now describe the identification of DEK as this 43-kDa pets factor. Using a modified Southwestern screening procedure, we find that DEK can recognize the pets element. We demonstrate the ability of recombinant DEK to bind specifically to the pets site using the electrophoretic mobility shift assay (EMSA) and DNase I footprinting. "Supershift" EMSA further confirms that DEK is the dominant protein binding to the pets site in T cell extracts. Our findings show that DEK is a site-specific DNA binding protein that is likely involved in transcriptional regulation and signal transduction. This has implications for multiple pathogenic processes, including hematologic malignancies, arthritis, ataxia-telangiectasia, and AIDS caused by HIV-2.
dek癌基因的产物是43 kDa的DEK核蛋白。DEK最初是在急性髓性白血病的一种特定亚型中与CAN核孔蛋白融合时被鉴定出来的。DEK也已被证明是少关节起病的幼年类风湿性关节炎患者的自身抗原。此外,DEK的最后65个氨基酸可以部分逆转共济失调毛细血管扩张症患者细胞的易突变表型。然而,尽管有这些显著的疾病关联,DEK的功能仍不清楚。HIV-2周边增强子元件(pets)是在HIV-2增强子中两个Elf-1结合位点之间发现的富含TG的元件。无论增强子是单独由佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)、单独由植物血凝素(PHA)、PMA加PHA、针对T细胞受体的可溶性抗体、针对T细胞受体的固定化抗体还是由抗原刺激,pets元件都能介导转录激活。以前,我们纯化并鉴定了pets因子,证明它是一种43 kDa的核蛋白。我们现在描述将DEK鉴定为这种