人CREB4转核机制的初步研究

cAMP 反应元件结合蛋白（cAMP response element-binding proteins，CREB）是一个哺乳动物转录因子家族，通过cAMP 反应元件（cAMP response element，CRE）调节cAMP和钙离子依赖性基因的表达.CREB4是CREB转录因子家族的一员.经酵母双杂交筛选人胎脑文库发现CREB4^215-279aa可能与核转运因子kayopherinα2相互作用，提示karyopherinα2可能参与CREB4的跨膜转运过程.亚细胞定位结果显示，CREB4全长定位于细胞质，而缺失C端假定转膜结构域的CREB4^1-279aa蛋白则转移至细胞核内.荧光共定位进一步显示，CREB4和karyopherinα2共定位于细胞质中，CREB4^1-279aa和karyopherinα2共定位于细胞核中.结果提示C端被切除之后，CREB4被karyopherinα2转运到核内发挥转录作用.     

Cyclic AMP response element binding protein (CREB) participates in the heat-inducible expression of humanhsp90β gene

Human heat shock protein 90b gene ( hsp90b ) is a constitutively expressed heat shock gene existing in most of cell types tested that can be further induced by heat shock. Chloramphenical acetyl transferase (CAT) reporter plasmids driven by different regulatory fragments of hsp90b gene were constructed and transfected into Jurkat cells to explore the role of a cAMP response element (CRE) in the upstream of the gene. Results show that, in comparison with the wild type construct, a severe reduction (~2/3) in the increased folds of promoter activity induced by heat shock at 42℃ for 1 h was observed in a construct with CRE-containing fragment (-173/-91bp) deleted. Electrophoretic mobility shift assays (EMSA) showed that phosphorylated CRE-binding protein (CREB) in the nuclear extract of heat shocked Jurkat cells is specifically bound to the fragment. Additionally, both of the phosphorylation on CREB and the activity of protein kinase A (PKA) were found in Jurkat cells to be enhanced with extending time of heat shock treatment. Our results indicate that in addition to the intronic HSE/HSF pathway, phosphorylated CREB also participates in the heat shock induced expression of human hsp90b gene via its interaction with CRE which may be regulated by PKA-sig- naling pathway.     

CREB regulates hepatic gluconeogenesis through the coactivator PGC-1

When mammals fast, glucose homeostasis is achieved by triggering expression of gluconeogenic genes in response to glucagon and glucocorticoids. The pathways act synergistically to induce gluconeogenesis (glucose synthesis), although the underlying mechanism has not been determined. Here we show that mice carrying a targeted disruption of the cyclic AMP (cAMP) response element binding (CREB) protein gene, or overexpressing a dominant-negative CREB inhibitor, exhibit fasting hypoglycaemia [corrected] and reduced expression of gluconeogenic enzymes. CREB was found to induce expression of the gluconeogenic programme through the nuclear receptor coactivator PGC-1, which is shown here to be a direct target for CREB regulation in vivo. Overexpression of PGC-1 in CREB-deficient mice restored glucose homeostasis and rescued expression of gluconeogenic genes. In transient assays, PGC-1 potentiated glucocorticoid induction of the gene for phosphoenolpyruvate carboxykinase (PEPCK), the rate-limiting enzyme in gluconeogenesis. PGC-1 promotes cooperativity between cyclic AMP and glucocorticoid signalling pathways during hepatic gluconeogenesis. Fasting hyperglycaemia is strongly correlated with type II diabetes, so our results suggest that the activation of PGC-1 by CREB in liver contributes importantly to the pathogenesis of this disease.     

化学防癌药物keap1-nrf2信号通路活化剂的研究进展

化学防癌是应用天然的或合成的化学物质来阻断、抑制或治疗癌症的过程,这一过程与肝内的Ⅱ相代谢酶的作用有关.研究表明,Keap1/Nrf2/ARE信号传导通路能有效地诱导Ⅱ相代谢酶基因的表达,而这一作用是作为碱性亮氨酸拉链蛋白家族的转录因子nrf2从胞质蛋白keap1上解离,然后转位至核内并与ARE结合后完成.这一传导通路机制加深了我们对于化学防癌机制的认识,并且为化学防癌药物的筛选提供了有效的靶点.就以Keap1/Nrf2/ARE传导通路为靶点的化学防癌药物的国内外研究进展进行了综述.