Wilkinson D G, Wilson J, Hames B D
Biochem J. 1983 Dec 15;216(3):567-74. doi: 10.1042/bj2160567.
The pattern of synthesis of the spore coat proteins during development of Dictyostelium discoideum has been determined by using immunoprecipitation with spore protein antibody. SP170, SP103, 'SP94', SP82, SP76 and SP55 are all first synthesized just prior to the 'Mexican hat' stage of development (16-18h), but the synthesis of SP72 is delayed. This protein is apparently synthesized as a precursor, P66, which is modified during spore maturation to yield SP72. The nature of the modification is unknown. At their peak period of synthesis during early culmination (18-20h), the spore coat proteins account for 5-9% of total protein synthesis. Shortly after synthesis, these proteins are inserted into the spore coat, where all except SP103 become disulphide-cross-linked during the period 24-30h. SP3 does not accumulate until disulphide-cross-linking of the major spore coat proteins occurs and is itself disulphide-cross-linked into the spore coat. Several additional proteins that are accumulated during development have also been identified, namely P31, P25, P21 and P18. P25 first appears at 18-20h and then continues to be made throughout development. P31 synthesis begins at 12-14h and then largely ceases after approx. 20 h of development. The genes for both P21 and P18 are first expressed early in development, starting at 9-12h. P21 synthesis ceases at approx. 14h, but P18 continues to be synthesized throughout the rest of development. The marked differences in the time period of accumulation of these proteins compared with the co-ordinated syntheses of SP170, SP103, 'SP94', SP82, SP76 and SP55 provide a useful system for analysis of the mechanism of temporal gene expression during development.
通过使用孢子蛋白抗体进行免疫沉淀,已确定了盘基网柄菌发育过程中孢子壁蛋白的合成模式。SP170、SP103、“SP94”、SP82、SP76和SP55均在发育的“墨西哥帽”阶段(16 - 18小时)之前首次合成,但SP72的合成延迟。这种蛋白质显然以前体P66的形式合成,在孢子成熟过程中经过修饰产生SP72。修饰的性质尚不清楚。在早期 culmination(18 - 20小时)的合成高峰期,孢子壁蛋白占总蛋白合成的5 - 9%。合成后不久,这些蛋白质被插入孢子壁,其中除SP103外,所有蛋白质在24 - 30小时期间形成二硫键交联。SP3直到主要孢子壁蛋白的二硫键交联发生后才积累,并自身二硫键交联到孢子壁中。在发育过程中积累的其他几种蛋白质也已被鉴定出来,即P31、P25、P21和P18。P25在18 - 20小时首次出现,然后在整个发育过程中持续合成。P31的合成在12 - 14小时开始,然后在发育约20小时后基本停止。P21和P18的基因在发育早期(9 - 12小时)开始首次表达。P21的合成在约14小时停止,但P18在发育的其余阶段继续合成。与SP170、SP103、“SP94”、SP82、SP76和SP55的协调合成相比,这些蛋白质积累时间段的显著差异为分析发育过程中时间基因表达的机制提供了一个有用的系统。