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通过补骨脂素光反应检测兔网织红细胞80S核糖体-mRNA复合物中未加帽流感病毒体外转录本与18S rRNA之间的碱基对形成。

Base-pair formation between noncapped influenza virus in vitro transcripts and 18S rRNA in rabbit reticulocyte 80S ribosome-mRNA complexes detected by psoralen photoreaction.

作者信息

Tanaka M, Hibasami H, Nagai J, Nakashima K

出版信息

J Biochem. 1984 Jan;95(1):125-30. doi: 10.1093/oxfordjournals.jbchem.a134574.

DOI:10.1093/oxfordjournals.jbchem.a134574
PMID:6706901
Abstract

RNA-RNA interactions between 18S ribosomal RNA and noncapped influenza cRNA were detected by psoralen photochemical cross-linking in reticulocyte 80S ribosome-cRNA complexes. In vitro transcripts of type A influenza virus synthesized by endogenous RNA polymerase with adenylyl-(3'----5')-guanosine primer formed 80S complexes with rabbit reticulocyte ribosomes. The extent of the complex formation by these noncapped cRNAs was less than that by the m7G-capped reovirus in vitro transcripts, but the former RNAs in the 80S complexes were cross-linked to 18S rRNA as efficiently as the latter RNAs by photoreaction with an RNA cross-linking agent, 4'-aminomethyl-4,5',8-trimethylpsoralen. These results suggested that mRNA with or without the cap structure on the 5'-terminal can form complexes with the ribosomes in a eukaryotic cell-free translation system by base-pair formation with 18S rRNA. Correspondingly, sequences capable of forming extensive base-pairs including four- to five-base complementarities were found between the 3'-terminal of rabbit reticulocyte 18S ribosomal RNA and the 5'-noncoding regions of either influenza virus transcripts or reovirus mRNA.

摘要

通过补骨脂素光化学交联法在网织红细胞80S核糖体-cRNA复合物中检测到18S核糖体RNA与无帽流感病毒cRNA之间的RNA-RNA相互作用。由内源性RNA聚合酶与腺苷酰-(3'→5')-鸟苷引物合成的甲型流感病毒体外转录本与兔网织红细胞核糖体形成80S复合物。这些无帽cRNA形成复合物的程度低于7-甲基鸟苷帽化的呼肠孤病毒体外转录本,但80S复合物中的前一种RNA通过与RNA交联剂4'-氨甲基-4,5',8-三甲基补骨脂素的光反应与18S rRNA交联的效率与后一种RNA相同。这些结果表明,5'-末端有或无帽结构的mRNA在真核无细胞翻译系统中可通过与18S rRNA形成碱基对与核糖体形成复合物。相应地,在兔网织红细胞18S核糖体RNA的3'-末端与流感病毒转录本或呼肠孤病毒mRNA的5'-非编码区之间发现了能够形成包括四到五个碱基互补的广泛碱基对的序列。

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