Soderberg L S, Tewari R P, Solotorovsky M
Infect Immun. 1976 Jun;13(6):1531-8. doi: 10.1128/iai.13.6.1531-1538.1976.
Macrophage ribonucleic acid (RNA) synthesis is an important metabolic process intimately related to the function of these cells. Mouse peritoneal macrophage RNA was extracted with phenol in the presence of bentonite and electrophoresed on composite agarose-polyacrylamide gels. The pulse-chase technique was used to follow the precursor relationships in macrophage ribosomal RNA (rRNA) maturation. The rRNA species at 18S and 28S appeared at 15 and 45 min, respectively, after RNA synthesis was halted. Their appearance corresponded closely to decreases in the rRNA precursors at 45S, 36S, and 34S. Studies of RNA methylation aided in confirming the identity of these ribosomal species. Unmethylated RNA species appeared as messenger RNA between 5S and 15S, and at about 55S probably represented heterodisperse nuclear RNA. When normal macrophages were incubated with heat-killed Salmonella enteritidis, an acceleration in the maturation of RNA was observed. The accelerated maturation was indicated by the earlier appearance of 28S rRNA and the more rapid development of an equilibrium state, where further labeling did not change the RNA profile. In macrophage RNA from mice immunized with S. enteritidis, rRNA species appeared rapidly but did not accumulate to the same extent as observed for normal macrophages. Precursor rRNA and other RNA species developed as usual, suggesting specific degradation of mature rRNA. Such rRNA wastage could indicate a mechanism controlling ribosome assembly in the non-proliferating activated macrophage. The pattern of RNA synthesis in immune macrophages was essentially unchanged by the presence of heat-killed S. enteritidis in vitro.
巨噬细胞核糖核酸(RNA)合成是一个与这些细胞功能密切相关的重要代谢过程。在膨润土存在的情况下,用苯酚提取小鼠腹腔巨噬细胞RNA,并在复合琼脂糖-聚丙烯酰胺凝胶上进行电泳。采用脉冲追踪技术来追踪巨噬细胞核糖体RNA(rRNA)成熟过程中的前体关系。在RNA合成停止后,18S和28S的rRNA种类分别在15分钟和45分钟出现。它们的出现与45S、36S和34S的rRNA前体的减少密切相关。RNA甲基化研究有助于确认这些核糖体种类的身份。未甲基化的RNA种类在5S至15S之间表现为信使RNA,而在约55S处可能代表异质核RNA。当正常巨噬细胞与热灭活的肠炎沙门氏菌一起孵育时,观察到RNA成熟加速。28S rRNA的较早出现以及平衡状态的更快发展表明了成熟加速,在这种平衡状态下进一步标记不会改变RNA图谱。在用肠炎沙门氏菌免疫的小鼠的巨噬细胞RNA中,rRNA种类迅速出现,但积累程度不如正常巨噬细胞。前体rRNA和其他RNA种类照常发展,表明成熟rRNA发生了特异性降解。这种rRNA损耗可能表明在非增殖性活化巨噬细胞中控制核糖体组装的一种机制。体外存在热灭活的肠炎沙门氏菌时,免疫巨噬细胞中的RNA合成模式基本不变。