Body B A, Brownstein B H
J Bacteriol. 1976 Jun;126(3):1149-55. doi: 10.1128/jb.126.3.1149-1155.1976.
Pulse-labeled cells of Bacillus megaterium were converted to protoplasts, and lysates of the protoplasts were analyzed by sucrose gradient sedimentation. Precursor ribonucleoprotein (RNP) particles then appeared predominantly as 50S and 30S precursor ribosomal subunits. Polyacrylamide gel electrophoresis of the ribosomal ribonucleic acid from the 50S and 30S RNP particles confirmed their precursor nature since they were shown to contain precursor 23S and 16S ribosomal ribonucleic acid, respectively. Treatment of protoplast lysates with 0.5% deoxycholate prior to sedimentation analysis resulted in a markedly different radioactivity profile. The 50S RNP particles were no longer present, but 43S particles were observed in addition to increased amounts of pulse-labeled material sedimenting at 30S and slower. Extracts from cells broken in a French press showed a profile from sucrose gradient sedimentation similar to that of the deoxycholate-treated protoplast lysate. These data suggest that the nature of the precursor ribosomal particles appears to be a function of the method of cell disruption or detergent treatment of the cell extract preparation. The observed 50S and 30S RNP particles may be the major precursor ribosomal subunits in vivo; the slower-sedimenting species could result from some form of breakdown or change in the configuration of the 50S and 30S precursors.
对巨大芽孢杆菌进行脉冲标记的细胞转化为原生质体,并通过蔗糖梯度沉降分析原生质体的裂解物。前体核糖核蛋白(RNP)颗粒主要表现为50S和30S前体核糖体亚基。对来自50S和30S RNP颗粒的核糖体核糖核酸进行聚丙烯酰胺凝胶电泳,证实了它们的前体性质,因为它们分别被证明含有前体23S和16S核糖体核糖核酸。在沉降分析之前,用0.5%脱氧胆酸盐处理原生质体裂解物,导致放射性分布明显不同。50S RNP颗粒不再存在,但除了在30S及更慢沉降速度下脉冲标记物质的量增加外,还观察到了43S颗粒。用法国压榨机破碎细胞得到的提取物,其蔗糖梯度沉降图谱与经脱氧胆酸盐处理的原生质体裂解物相似。这些数据表明,前体核糖体颗粒的性质似乎是细胞破碎方法或细胞提取物制备过程中去污剂处理方式的函数。观察到的50S和30S RNP颗粒可能是体内主要的前体核糖体亚基;沉降较慢的物种可能是50S和30S前体某种形式的分解或构型变化导致的。