Max-Planck-Institut für Biochemie, D-8033 Martinsried, FRG.
EMBO J. 1986 Dec 20;5(13):3437-40. doi: 10.1002/j.1460-2075.1986.tb04666.x.
When vegetative cells of Dictyostelium discoideum were incubated in nutrient medium containing 5 mM caffeine, adenylate cyclase was induced and exceeded control values 30-fold after 6 h of incubation. The effect was dose-dependent and blocked by cycloheximide. Contact site A, a developmentally regulated membrane glycoprotein characteristic of aggregation-competent cells, was not induced by the treatment. The expression of a developmentally regulated gene coding for an unknown protein and hybridizing with the cDNA clone P26E8 was also stimulated, indicating that the regulation of expression occurs at the transcript level. Cells pretreated with caffeine in growth medium, washed and developed in starvation buffer showed an acceleration of development by 2 h as judged by stream formation and the appearance of contact sites A. The same effects were observed when cells were incubated under heat shock conditions (30 degrees C). The results indicate that caffeine stimulates the expression of devevelopmentally regulated early genes, and that their products, together with exogeneous factor(s), initiate subsequent steps of development.
在用含有 5mM 咖啡因的营养培养基孵育的 Dictyostelium discoideum 营养细胞中,腺嘌呤环化酶被诱导,孵育 6 小时后,其活性超过对照值 30 倍。该效应呈剂量依赖性,并被环己亚胺所阻断。接触位点 A 是一种与聚集能力细胞相关的发育调节膜糖蛋白,该处理并未诱导其产生。一种发育调节基因的表达也被刺激,该基因编码一种未知蛋白并与 cDNA 克隆 P26E8 杂交,这表明表达的调控发生在转录水平。用生长培养基中咖啡因预处理的细胞,经洗涤并在饥饿缓冲液中发育,根据流形成和接触位点 A 的出现,其发育速度加快了 2 小时。当细胞在热休克条件(30°C)下孵育时,也观察到相同的效果。结果表明,咖啡因刺激发育调节早期基因的表达,其产物与外源因子一起启动随后的发育步骤。