NON-IONIC OLIGONUCLEOSIDE METHYLPHOSPHONATES .4. SELECTIVE-INHIBITION OF ESCHERICHIA-COLI PROTEIN-SYNTHESIS AND GROWTH BY NON-IONIC OLIGONUCLEOTIDES COMPLEMENTARY TO THE 3' END OF 16S RIBOSOMAL-RNA
NON-IONIC OLIGONUCLEOSIDE METHYLPHOSPHONATES .4. SELECTIVE-INHIBITION OF ESCHERICHIA-COLI PROTEIN-SYNTHESIS AND GROWTH BY NON-IONIC OLIGONUCLEOTIDES COMPLEMENTARY TO THE 3' END OF 16S RIBOSOMAL-RNA
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DOI：
10.1073/pnas.78.3.1537
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
TSO, POP
中科院分区：
文献类型：
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作者：
JAYARAMAN, K;MCPARLAND, K;TSO, POP
合成了一系列非离子型寡核苷酸类似物，脱氧核糖核苷甲基膦酸酯。这些化合物的碱基序列d（ApGpGp）、d（ApGpGp）2和d[（ApGpGp）2 T]与在细菌16 S rRNA的3“末端发现的Shine-Dalgarno序列（-A-C-C-U-C-C-U-）互补。测试这些非离子寡核苷酸类似物在大肠杆菌无细胞系统中抑制mRNA体外翻译的能力。大肠杆菌和兔网织红细胞。在急诊coli系统中，d（ApGpGp）2和d[（ApGpGp）2 T]能有效抑制MS-2 RNA介导的蛋白质合成，但对poly（U）或poly（A）介导的多肽合成的影响很小。在网织红细胞系统中，这些化合物对珠蛋白mRNA的翻译没有显著影响。观察到d[（ApGpGp）2[3 H]T]与70 S核糖体结合（结合常数，2.0 × 1017）。104 M-1，37 °C）连同这些化合物对蛋白质合成的抑制作用的特异性一起强烈地表明，翻译的抑制是类似物与16 S rRNA的Shine-Dalgarno序列结合的结果。甲基膦酸单核苷酸抑制蛋白质合成（不同时抑制RNA合成）和大肠杆菌的集落形成。coliML 308-225（一种渗透性突变体），其细胞壁含有可忽略量的脂多糖，但对野生型E.大肠杆菌B。大肠杆菌摄取寡脱氧核苷甲基膦酸酯的初步结果。coli B中的结果表明，这些细胞对长于4个核苷酸单位的寡聚体是不可渗透的。尽管寡脱氧核糖核苷甲基膦酸酯被培养的哺乳动物细胞吸收，但这一系列类似物对转化的人细胞的集落形成具有可忽略的抑制作用。这类非离子寡核苷酸类似物可用于探测和调节活细胞内具有确定序列的核酸的功能和结构。
A series of nonionic oligonucleotide analogs, the deoxyribooligonucleoside methylphosphonates, were synthesized. The base sequences of these compounds, d(ApGpGp), d(ApGpGp)2 and d[(ApGpGp)2T], are complementary to the Shine-Dalgarno sequence (-A-C-C-U-C-C-U-) found at the 3'' end of bacterial 16S rRNA. These nonionic oligonucleotide analogs were tested for their ability to inhibit the in vitro translation of mRNA in cell-free systems of E. coli and rabbit reticulocyte. In the E. coli system, d(ApGpGp)2 and d[(ApGpGp)2T] effectively inhibited MS-2 RNA-directed protein synthesis but they had much less effect on poly(U)- or poly(A)-directed polypeptide synthesis. In the reticulocyte system, these compounds had no significant effect on the translation of globin mRNA. The observation that d[(ApGpGp)2[3H]T] binds to 70S ribosomes (association constant, 2.0 .times. 104 M-1, 37.degree. C) together with the specificity of the inhibitory action of these compounds on protein synthesis strongly suggests that inhibition of translation is a consequence of analog binding to the Shine-Dalgarno sequence of 16S rRNA. The oligonucleoside methylphosphonates inhibited protein synthesis (without concurrent inhibition of RNA synthesis) and colony formation by E. coli ML 308-225 (a permeable mutant) whose cell wall contains negligible quantities of lipopolysaccharide but had no effect on wild-type E. coli B. Preliminary results on the uptake of oligodeoxyribonucleoside methylphosphonates by E. coli B show that these cells are not permeable to oligomers longer than 4 nucleotidyl units. Although oligodeoxyribonucleoside methylphosphonates are taken up by mammalian cells in culture, this series of analogs had negligible inhibitory effects on colony formation by transformed human cells. This class of nonionic oligonucleotide analogs may be used to probe and regulate the function and structure of nucleic acids of defined sequence within living cells.