Zhang Jun, Yang Jinbo, Roy Sanjit K, Tininini Silvia, Hu Jiadi, Bromberg Jacqueline F, Poli Valeria, Stark George R, Kalvakolanu Dhananjaya V
Greenebaum Cancer Center, Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
Proc Natl Acad Sci U S A. 2003 Aug 5;100(16):9342-7. doi: 10.1073/pnas.1633516100. Epub 2003 Jul 16.
GRIM-19 (gene associated with retinoid-IFN-induced mortality 19), isolated as a cell death activator in a genetic screen used to define mechanisms involved in IFN-beta- and retinoic acid-induced cell death, codes for a approximately 16-kDa protein that induces apoptosis in a number of cell lines. Antisense ablation of GRIM-19 caused resistance to cell death induced by IFN plus retinoic acid and conferred a growth advantage to cells. To understand the molecular bases for its cell death regulatory activity, we used a yeast two-hybrid screen and identified that the transcription factor STAT3 (signal transducer and activator of transcription 3) binds to GRIM-19. GRIM-19 inhibits transcription driven by activation of STAT3, but not STAT1. It neither inhibits the ligand-induced activation of STAT3 nor blocks its ability to bind to DNA. Mutational analysis indicates that the transactivation domain of STAT3, especially residue S727, is required for GRIM-19 binding. Because GRIM-19 does not bind significantly to other STATs, our studies identify a specific inhibitor of STAT3. Because constitutively active STAT3 up-regulates antiapoptotic genes to promote tumor survival, its inhibition by GRIM-19 also demonstrates an antioncogenic effect exerted by biological therapeutics.
GRIM-19(类视黄醇-干扰素诱导死亡相关基因19),在用于确定β-干扰素和视黄酸诱导细胞死亡机制的基因筛选中作为细胞死亡激活剂被分离出来,编码一种约16 kDa的蛋白质,该蛋白质可在多种细胞系中诱导细胞凋亡。GRIM-19的反义缺失导致对干扰素加视黄酸诱导的细胞死亡产生抗性,并赋予细胞生长优势。为了解其细胞死亡调节活性的分子基础,我们利用酵母双杂交筛选,确定转录因子STAT3(信号转导和转录激活因子3)与GRIM-19结合。GRIM-19抑制由STAT3激活驱动的转录,但不抑制STAT1。它既不抑制配体诱导的STAT3激活,也不阻断其与DNA结合的能力。突变分析表明,STAT3的反式激活结构域,尤其是S727残基,是GRIM-19结合所必需的。由于GRIM-19与其他STATs无明显结合,我们的研究确定了一种STAT3的特异性抑制剂。由于组成型激活的STAT3上调抗凋亡基因以促进肿瘤存活,GRIM-19对其的抑制也证明了生物治疗剂发挥的抗癌作用。