Steel L F, Jacobson A
Mol Cell Biol. 1987 Mar;7(3):965-72. doi: 10.1128/mcb.7.3.965-972.1987.
Throughout the developmental program of Dictyostelium discoideum there are substantial changes in the rates of both ribosome utilization and rRNA transcription and processing. We examined the regulation of ribosomal protein (r-protein) gene expression and found that, at the start of development, expression of these genes was drastically and specifically reduced by a block to translational initiation. An apparently separate event signals a sudden decrease in the relative amount of r-protein mRNA at about 10 h of development, a time when aggregated amoebae are forming tight cell-cell contacts. For the first 9 h of development, the relative amount of r-protein mRNA remained essentially unchanged and comparable to levels detected in growing cells. While the r-protein mRNAs were almost fully loaded on polysomes during vegetative growth, they were specifically excluded from polysomes at the start of development. The translational block was not the result of irreversible structural changes which inactivate the r-protein mRNAs since they remained translatable both in vitro, in wheat germ extracts, and in vivo, where they were recruited onto polysomes in the presence of the elongation inhibitor cycloheximide. In addition, precise measurements of poly(A) tail lengths on individual hybrid-selected mRNA species showed that there is no difference in the poly(A) tail length of r-protein mRNA isolated from growing cells and 1-h developing cells. Therefore, changes in translational efficiency cannot be attributed to cleavage of poly(A) tails.
在盘基网柄菌的整个发育过程中,核糖体利用率以及rRNA转录和加工速率都发生了显著变化。我们研究了核糖体蛋白(r蛋白)基因表达的调控,发现发育开始时,这些基因的表达因翻译起始受阻而急剧且特异性地降低。一个明显不同的事件表明,在发育约10小时时,r蛋白mRNA的相对量突然下降,此时聚集的变形虫正在形成紧密的细胞间接触。在发育的前9小时,r蛋白mRNA的相对量基本保持不变,与在生长细胞中检测到的水平相当。虽然r蛋白mRNA在营养生长期间几乎完全负载在多聚核糖体上,但在发育开始时它们被特异性地排除在多聚核糖体之外。翻译受阻并非是由于不可逆的结构变化使r蛋白mRNA失活,因为它们在体外小麦胚提取物中以及在体内,即在存在延伸抑制剂环己酰亚胺的情况下被招募到多聚核糖体上时,仍然是可翻译的。此外,对单个杂交选择的mRNA种类上的聚(A)尾长度进行的精确测量表明,从生长细胞和发育1小时的细胞中分离出的r蛋白mRNA的聚(A)尾长度没有差异。因此,翻译效率的变化不能归因于聚(A)尾的切割。