Geisser M, Mackie G A
Eur J Biochem. 1976 Nov 1;70(1):159-70. doi: 10.1111/j.1432-1033.1976.tb10966.x.
Ribosomal protein S4 of Escherichia coli was bound to 16-S ribosomal RNAs from several bacterial species and the complexes digested with pancreatic ribonuclease in an effort to isolate heterologous RNA binding sites for protein S4. 16-S RNAs from Aeromonas punctata, Pseudomonas fluorescens and Vibrio cuneatus each gave rise to protected fragments whose electrophoretic mobility was 7S, i.e. similar to that of the fragment generated from E. coli 16-S RNA using the same conditions. No comparable fragment was obtained from 16-S RNA of either Bacillus subtilis or Bacillus stearothermophilus, if E. coli protein S4 was present prior to digestion. The protected 7-S RNA fragment from A. punctata and the subfragments obtained from it by gel electrophoresis under denaturing conditions were characterized further by fingerprinting and nucleotide sequence analysis. The sequence of many of the T1 ribonuclease oligonucleotides was obtained and compared to those from the E. coli 7-S fragment. This has permitted a tentative identification of the sequences of A. punctata 16-S RNA which are protected by E. coli protein S4, namely, the regions homologous to the E. coli sequence from section M through C''. The fingerprints of the protected 7-S fragments from both P. fluorescens and V. cuneatus were sufficiently different from that of the E. coli 7-S fragment that no conclusions regarding sequence homologies could be drawn.
将大肠杆菌的核糖体蛋白S4与几种细菌物种的16 - S核糖体RNA结合，并使用胰核糖核酸酶消化这些复合物，以分离蛋白S4的异源RNA结合位点。来自点状气单胞菌、荧光假单胞菌和楔形弧菌的16 - S RNA均产生了电泳迁移率为7S的受保护片段，即在相同条件下，其与从大肠杆菌16 - S RNA产生的片段的电泳迁移率相似。如果在消化前存在大肠杆菌蛋白S4，则枯草芽孢杆菌或嗜热脂肪芽孢杆菌的16 - S RNA均未获得可比片段。通过指纹图谱和核苷酸序列分析，进一步对来自点状气单胞菌的受保护7 - S RNA片段及其在变性条件下通过凝胶电泳获得的亚片段进行了表征。获得了许多T1核糖核酸酶寡核苷酸的序列，并将其与来自大肠杆菌7 - S片段的序列进行了比较。这使得能够初步鉴定点状气单胞菌16 - S RNA中受大肠杆菌蛋白S4保护的序列，即与从M区到C''区的大肠杆菌序列同源的区域。来自荧光假单胞菌和楔形弧菌的受保护7 - S片段的指纹图谱与大肠杆菌7 - S片段的指纹图谱差异足够大，以至于无法得出关于序列同源性的结论。
