Ma P C, Siu C H
Banting and Best Department of Medical Research, University of Toronto, Ontario, Canada.
Mol Cell Biol. 1990 Jul;10(7):3297-306. doi: 10.1128/mcb.10.7.3297-3306.1990.
The EDTA-resistant cell-cell adhesion expressed at the aggregation stage of Dictyostelium discoideum is mediated by a cell surface glycoprotein of Mr 80,000 (gp80). The expression of gp80 is developmentally regulated by cyclic AMP (cAMP). In vitro nuclear run-on experiments show that transcription of the gp80 gene is initiated soon after the onset of development. The basal level of gp80 transcription is significantly augmented by exogenous cAMP pulses. Interestingly, in analog studies, 2'-deoxy-cAMP, 8-bromo-cAMP, and N6-monobutyryl-cAMP are all capable of inducing a rapid accumulation of gp80 mRNA, suggesting the presence of a unique cAMP receptor that responds equally well to these analogs. To determine whether intracellular cAMP plays a role in the regulation of gp80 expression, caffeine was used to block cAMP-induced receptor-mediated adenylate cyclase activation. Expression of gp80 mRNA was blocked in caffeine-treated cells but could be substantially restored by treatment with exogenous cAMP pulses, suggesting that adenylate cyclase activation is not required. gp80 expression was also examined in the signal transduction mutants synag 7 and frigid A. In both mutants, gp80 was expressed at the basal level. Pulses of cAMP as well as 2'-deoxy-cAMP and N6-monobutyryl-cAMP were capable of restoring the normal level of gp80 expression in synag 7 cells. These results, taken together, indicate bimodal regulation of gp80 expression during development and the involvement of a novel cAMP receptor in the transmembrane signalling pathway that regulates gp80 gene expression.
盘基网柄菌聚集阶段表达的抗乙二胺四乙酸(EDTA)的细胞间黏附作用由一种分子量为80,000的细胞表面糖蛋白(gp80)介导。gp80的表达受环磷酸腺苷(cAMP)的发育调控。体外核转录实验表明,gp80基因的转录在发育开始后不久就启动。外源性cAMP脉冲显著增强了gp80转录的基础水平。有趣的是,在类似物研究中,2'-脱氧-cAMP、8-溴-cAMP和N6-单丁酰-cAMP都能够诱导gp80 mRNA的快速积累,这表明存在一种对这些类似物反应同样良好的独特cAMP受体。为了确定细胞内cAMP是否在gp80表达的调控中起作用,使用咖啡因来阻断cAMP诱导的受体介导的腺苷酸环化酶激活。在咖啡因处理的细胞中,gp80 mRNA的表达被阻断,但通过外源性cAMP脉冲处理可以基本恢复,这表明不需要腺苷酸环化酶激活。还在信号转导突变体synag 7和frigid A中检测了gp80的表达。在这两个突变体中,gp80都以基础水平表达。cAMP以及2'-脱氧-cAMP和N6-单丁酰-cAMP的脉冲能够恢复synag 7细胞中gp80表达的正常水平。综合这些结果,表明在发育过程中gp80表达存在双峰调控,并且在调节gp80基因表达的跨膜信号通路中涉及一种新型cAMP受体。