In vivo UV cross-linking of U snRNAs that participate in trypanosome trans-splicing.
In vivo UV cross-linking of U snRNAs that participate in trypanosome trans-splicing.
复制标题
参与锥虫转拼的 U snRNA 的体内紫外交联。
DOI：
10.1101/gad.5.10.1859
复制
发表时间：
1991
影响因子：
10.5
通讯作者：
Agabian,N
中科院分区：
文献类型：
--
作者：
Watkins,KP;Agabian,N
布氏锥虫中mRNA的成熟涉及反式剪接反应，由此称为剪接前导（SL）RNA的小RNA的5' 39个核苷酸与前mRNA转录物连接。反式剪接反应在机制上类似于核前mRNA的顺式剪接，并且该过程需要U2、U4和U6 snRNA的同源物。在这里介绍的工作中，潜在的RNA-RNA之间的相互作用的SL RNA和锥虫的U snRNA的紫外光诱导的RNA-RNA交联在体内检查。我们检测到U2和U6 RNA之间的交联，正如预期的那样，锥虫U4和U6 RNA之间的交联。后者包含广泛的序列互补性，被认为主要存在于单个RNP中。我们还检测到SL RNA物种在体内UV处理后，这可能代表SL RNA中的分子内交联或SL RNA与尚未鉴定的小RNA之间形成的交联。U2和U6 RNA之间的交联位置的作图与U2的5'结构域和U6 RNA的3'末端之间的碱基配对一致。这些结果表明，在体内，同源RNA-RNA相互作用的RNA同源物参与在锥虫的反式剪接和其他真核生物的顺式剪接。
The maturation of mRNAs in Trypanosoma brucei involves a trans-splicing reaction whereby the 5' 39 nucleotides of a small RNA, called the spliced leader (SL) RNA, are joined with a pre-mRNA transcript. The trans-splicing reaction appears mechanistically similar to cis-splicing of nuclear pre-mRNAs, and homologs of the U2, U4, and U6 snRNAs are required for the process. In the work presented here, potential RNA-RNA interactions between the SL RNA and the U snRNAs of trypanosomes were examined by UV light induction of RNA-RNA cross-links in vivo. We detected cross-linkage between U2 and U6 RNAs and, as might be expected, between the trypanosome U4 and U6 RNAs. The latter contain extensive sequence complementarity and are thought to exist predominantly in a single RNP. We also detected an SL RNA species following in vivo UV treatment, which may represent either an intramolecular cross-link in the SL RNA or a cross-link formed between the SL RNA and an as yet unidentified small RNA. Mapping of the cross-link position between U2 and U6 RNAs is consistent with base-pairing between the 5' domain of U2 and the 3' end of U6 RNA. These results reveal the existence, in vivo, of cognate RNA-RNA interactions in the RNA homologs that participate in trans-splicing in trypanosomes and cis-splicing in other eukaryotes.