用在分子茎内碱基配对存在干扰的人工5S RNA分子重建具有生物活性的50S核糖体亚基。
Reconstitution of biologically active 50S ribosomal subunits with artificial 5S RNA molecules carrying disturbances in the base pairing within the molecular stalk.
作者信息
Raué H A, Lorenz S, Erdmann V A, Planta R J
出版信息
Nucleic Acids Res. 1981 Mar 11;9(5):1263-9. doi: 10.1093/nar/9.5.1263.
Bacillus stearothermophilus 50S ribosomal subunits were reconstituted in vitro using artificial 5S RNA molecules constructed by combining parts of major and minor type (Raué et al. (1976) Europ. J. Biochem. 68, 169-176) B. licheniformis 5S RNA. The artificial 5S RNA molecules carry defined disturbances (A.C juxtapositions and extra G.U pairs) in the base pairing between the 5'- and 3'-terminal sequences of the molecule (the molecular stalk region). The biological activity of the reconstituted subunits was determined in an E. coli cell-free system programmed with poly-U. The results show that conservation of the base pairing within the molecular stalk is not required for biological activity of 5S RNA. Disturbances of the base pairing within this region do reduce the rate of reconstitution, however. Normal base pairing in the molecular stalk is thus required to ensure efficient ribosome assembly.
嗜热脂肪芽孢杆菌50S核糖体亚基在体外使用人工5S RNA分子进行重建，这些人工5S RNA分子是通过将地衣芽孢杆菌主要型和次要型5S RNA的部分序列组合构建而成（劳埃等人，（1976年）《欧洲生物化学杂志》68卷，第169 - 176页）。人工5S RNA分子在分子的5'和3'末端序列之间的碱基配对（分子茎区）中携带特定的干扰（A.C并列和额外的G.U对）。在以聚尿嘧啶编程的大肠杆菌无细胞系统中测定重建亚基的生物学活性。结果表明，5S RNA的生物学活性并不需要分子茎内碱基配对的保守性。然而，该区域内碱基配对的干扰确实会降低重建速率。因此，分子茎中的正常碱基配对是确保核糖体高效组装所必需的。
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