Ko Lan, Cardona Guemalli R, Henrion-Caude Alexandra, Chin William W
Department of Gene Regulation, Bone and Inflammation Research, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285, USA.
Mol Cell Biol. 2002 Jan;22(1):357-69. doi: 10.1128/MCB.22.1.357-369.2002.
Gene activation mediated by nuclear receptors is regulated in a tissue-specific manner and requires interactions between nuclear receptors and their cofactors. Here, we identified and characterized a tissue-specific coactivator, GT198, that interacts with the DNA-binding domains of nuclear receptors. GT198 was originally described as a genomic transcript that mapped to the human breast cancer susceptibility locus 17q12-q21 with unknown function. We show that GT198 exhibits a tissue-specific expression pattern in which its mRNA is elevated in testis, spleen, thymus, pituitary cells, and several cancer cell lines. GT198 is a 217-amino-acid nuclear protein that contains a leucine zipper required for its dimerization. In vitro binding and yeast two-hybrid assays indicated that GT198 interacted with nuclear receptors through their DNA-binding domains. GT198 potently stimulated transcription mediated by estrogen receptor alpha and beta, thyroid hormone receptor beta1, androgen receptor, glucocorticoid receptor, and progesterone receptor. However, the action of GT198 was distinguishable from that of the ligand-binding domain-interacting nuclear receptor coactivators, such as TRBP, CBP, and SRC-1, with respect to basal activation and hormone sensitivity. Furthermore, protein kinase A, protein kinase C, and mitogen-activated protein kinase can phosphorylate GT198 in vitro, and cotransfection of these kinases regulated the transcriptional activity of GT198. These data suggest that GT198 is a tissue-specific, kinase-regulated nuclear receptor coactivator that interacts with the DNA-binding domains of nuclear receptors.
由核受体介导的基因激活以组织特异性方式受到调控,并且需要核受体与其辅因子之间的相互作用。在此,我们鉴定并表征了一种组织特异性共激活因子GT198,它与核受体的DNA结合结构域相互作用。GT198最初被描述为一种基因组转录本,定位于人乳腺癌易感基因座17q12 - q21,功能未知。我们发现GT198呈现出组织特异性表达模式,其mRNA在睾丸、脾脏、胸腺、垂体细胞以及几种癌细胞系中升高。GT198是一种含有217个氨基酸的核蛋白,包含其二聚化所需的亮氨酸拉链。体外结合和酵母双杂交试验表明,GT198通过核受体的DNA结合结构域与它们相互作用。GT198能有效刺激由雌激素受体α和β、甲状腺激素受体β1、雄激素受体、糖皮质激素受体和孕激素受体介导的转录。然而,就基础激活和激素敏感性而言,GT198的作用与配体结合结构域相互作用的核受体共激活因子如TRBP、CBP和SRC - 1不同。此外,蛋白激酶A、蛋白激酶C和丝裂原活化蛋白激酶在体外可使GT198磷酸化,这些激酶的共转染调节了GT198的转录活性。这些数据表明,GT198是一种组织特异性、激酶调节的核受体共激活因子,它与核受体的DNA结合结构域相互作用。