...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Nucleic Acids Research >A NEW TECHNIQUE FOR THE CHARACTERIZATION OF LONG-RANGE TERTIARY CONTACTS IN LARGE RNA MOLECULES - INSERTION OF A PHOTOLABEL AT A SELECTED POSITION IN 16S RRNA WITHIN THE ESCHERICHIA COLI RIBOSOME
【24h】

A NEW TECHNIQUE FOR THE CHARACTERIZATION OF LONG-RANGE TERTIARY CONTACTS IN LARGE RNA MOLECULES - INSERTION OF A PHOTOLABEL AT A SELECTED POSITION IN 16S RRNA WITHIN THE ESCHERICHIA COLI RIBOSOME

机译：表征大RNA分子长链接触的新技术-在大肠杆菌核糖体中在16S RRNA的选定位置插入光标记。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

A new approach for inserting a photo-label at a selected position within the long ribosomal RNA molecules has been developed. The Escherichia coli 16S rRNA was cleaved at a single internucleotide bond, 1141-1142, with RNase H in the presence of a complementary chimeric oligonucleotide. 4-Thiouridine 5', 3'-diphosphate was ligated to the 3'-end of the 5' fragment at the cleavage site with T4 RNA ligase. The 16S rRNA fragments containing this added photo-reactive nucleotide were assembled together with total 30S ribosomal proteins into small ribosomal subunits. The ability of such 30S particles containing fragmented rRNA to form 70S ribosomes has been demonstrated previously. Crosslinks were induced within the 30S subunits by mild UV irradiation. The sites of crosslinking within the 16S rRNA were then analyzed using RNase H digestion and reverse transcription. Two crosslinks from the thio-nucleotide attached to nt C1141 of 16S rRNA were observed, namely to nt U1295 and G1272. These results are in agreement with the established proximity of helix 39 and 41 in the 3D structure of the 30S ribosomal subunit, as shown by other intra RNA crosslinking data. These data furthermore allow us to refine the structural arrangement of helices 41 and 39 relative to one another.
机译：已开发出一种在长核糖体RNA分子内选定位置插入光标记的新方法。在互补嵌合寡核苷酸的存在下，用RNase H在单核苷酸间键1141-1142切割大肠杆菌16S rRNA。用T4 RNA连接酶将4-硫代吡啶5'，3'-二磷酸连接到5'片段的3'末端的切割位点。包含该添加的光反应性核苷酸的16S rRNA片段与总的30S核糖体蛋白一起组装成小的核糖体亚基。先前已经证明了这种包含片段化的rRNA的30S颗粒形成70S核糖体的能力。通过温和的紫外线照射，在30S亚基内诱导了交联。然后使用RNase H消化和逆转录分析16S rRNA中的交联位点。观察到来自硫核苷酸的两个交联，其与16S rRNA的nt C1141相连，即与nt U1295和G1272相连。这些结果与30S核糖体亚基的3D结构中螺旋线39和41的确定接近度一致，如其他内部RNA交联数据所示。这些数据还使我们能够改进螺旋41和39相对于彼此的结构布置。

著录项

相似文献

  • 外文文献
  • 中文文献
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号