Evidence for a tRNA/rRNA interaction site within the peptidyltransferase center of the Escherichia coli ribosome.
Evidence for a tRNA/rRNA interaction site within the peptidyltransferase center of the Escherichia coli ribosome.
复制标题
大肠杆菌核糖体肽基转移酶中心内存在 tRNA/rRNA 相互作用位点的证据。
DOI：
10.1021/bi00428a073
复制
发表时间：
1989
期刊：
影响因子：
2.9
通讯作者：
Hill,WE
中科院分区：
文献类型：
--
作者：
Marconi,RT;Hill,WE
蒙大拿大学化学系，Missoula，Montana 59812 1988年2月26日接收; 1988年7月18日修订的Mandarin pt接收摘要：与23 S rRNA的碱基2497-2505互补的九碱基寡脱氧核糖核苷酸与50 S亚基和70 S核糖体杂交。通过硝酸纤维素过滤和蔗糖梯度离心技术测定探针与核糖体或核糖体亚基的结合。通过用核糖核酸酶H消化rRNA/cDNA异源双链体并对消化产物进行凝胶电泳，然后对较小的消化片段进行分离和测序来确定杂交位点的位置。发现cDNA探针与其rRNA靶位点特异性相互作用。研究了结合脱酰tRNAPhe对探针与50 S和70 S核糖体杂交的影响。脱酰tRNAphe的结合，无论有或没有添加聚（尿苷酸），引起探针结合50 S和70 S核糖体的衰减。在50 S亚基和70 S核糖体中，探针与23 SrRNA的杂交均降低约75%。这些结果表明，2497-2505位点内的碱基可能参与脱酰化的tRNA/rRNA相互作用。
Department of Chemistry, University of Montana, Missoula, Montana 59812 Received February 26, 1988; Revised Manuscript Received July 18, 1988 abstract: A nine-base oligodeoxyribonucleotide complementary to bases 2497-2505 of 23S rRNA was hybridized to both 50S subunits and 70S ribosomes. The binding of the probe to the ribosome or ribosomal subunits was assayed by nitrocellulose filtration and by sucrose gradient centrifugation techniques. The location of the hybridization site was determined by digestion of the rRNA/cDNA heteroduplex with ribonuclease H and gel electrophoresis of the digestion products, followed by the isolation and sequencing of the smaller digestion fragment. The cDNA probe was found to interact specifically with its rRNA target site. The effects on probe hybridization to both 50S and 70S ribosomes as a result of binding deacylated tRNAPhe were investigated. The binding of deacylated tRNAphe, eitherwith or without the addition of poly (uridylic acid), caused attenuation of probe binding to both 50S and 70S ribosomes. Probe hybridization to 23S rRNA was decreased byabout 75% in both 50S subunits and 70S ribosomes. These results suggest that bases within the 2497-2505 site may participate in a deacylated tRNA/rRNA interaction.