Yamashita H, Takenoshita M, Sakurai M, Bruick R K, Henzel W J, Shillinglaw W, Arnot D, Uyeda K
Dallas Veterans Affairs Medical Center and Department of Biochemistry, University of Texas Southwestern Medical Center, 4500 South Lancaster Road, Dallas, TX 75216, USA.
Proc Natl Acad Sci U S A. 2001 Jul 31;98(16):9116-21. doi: 10.1073/pnas.161284298. Epub 2001 Jul 24.
Carbohydrates mediate their conversion to triglycerides in the liver by promoting both rapid posttranslational activation of rate-limiting glycolytic and lipogenic enzymes and transcriptional induction of the genes encoding many of these same enzymes. The mechanism by which elevated carbohydrate levels affect transcription of these genes remains unknown. Here we report the purification and identification of a transcription factor that recognizes the carbohydrate response element (ChRE) within the promoter of the L-type pyruvate kinase (LPK) gene. The DNA-binding activity of this ChRE-binding protein (ChREBP) in rat livers is specifically induced by a high carbohydrate diet. ChREBP's DNA-binding specificity in vitro precisely correlates with promoter activity in vivo. Furthermore, forced ChREBP overexpression in primary hepatocytes activates transcription from the L-type Pyruvate kinase promoter in response to high glucose levels. The DNA-binding activity of ChREBP can be modulated in vitro by means of changes in its phosphorylation state, suggesting a possible mode of glucose-responsive regulation. ChREBP is likely critical for the optimal long-term storage of excess carbohydrates as fats, and may contribute to the imbalance between nutrient utilization and storage characteristic of obesity.
碳水化合物在肝脏中通过促进限速糖酵解酶和脂肪生成酶的快速翻译后激活以及编码许多这些相同酶的基因的转录诱导,介导其向甘油三酯的转化。碳水化合物水平升高影响这些基因转录的机制仍然未知。在这里,我们报告了一种转录因子的纯化和鉴定,该转录因子识别L型丙酮酸激酶(LPK)基因启动子内的碳水化合物反应元件(ChRE)。大鼠肝脏中这种ChRE结合蛋白(ChREBP)的DNA结合活性由高碳水化合物饮食特异性诱导。ChREBP在体外的DNA结合特异性与体内启动子活性精确相关。此外,在原代肝细胞中强制过表达ChREBP会响应高葡萄糖水平激活L型丙酮酸激酶启动子的转录。ChREBP的DNA结合活性可以在体外通过其磷酸化状态的变化进行调节,这提示了一种可能的葡萄糖反应调节模式。ChREBP可能对于将过量碳水化合物最佳地长期储存为脂肪至关重要,并且可能导致肥胖所特有的营养利用与储存之间的失衡。