A prokaryotic-like mode of cytoplasmic eukaryotic ribosome binding to the initiation codon during internal translation initiation of hepatitis C and classical swine fever virus RNAs
A prokaryotic-like mode of cytoplasmic eukaryotic ribosome binding to the initiation codon during internal translation initiation of hepatitis C and classical swine fever virus RNAs
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DOI：
10.1101/gad.12.1.67
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发表时间：
1998-01-01
影响因子：
10.5
通讯作者：
Hellen, CUT
中科院分区：
文献类型：
--
作者：
Pestova, TV;Shatsky, IN;Hellen, CUT
丙型肝炎病毒和猪瘟病毒mRNA翻译的起始是由内部核糖体进入引起的。我们重建内部核糖体进入体外纯化的翻译组件和监测组装的48 S核糖体preinitiation复合物的脚趾打印。核糖体亚基（40 S）在两种mRNA上形成稳定的二元复合物。这些RNA的复杂结构决定了起始密码子在二元复合物中核糖体“P”位点的正确定位。核糖体结合和定位在这些mRNA上不需要起始因子eIF 3、eIF 4A、eIF 4 B和eIF 4F，并且这些mRNA的翻译不受反式显性eIF 4A突变体的抑制。在这些二元核糖体复合物中加入Met-tRNA（i）（Met）、eIF 2和GTP，导致形成48 S前起始复合物。这种真核生物的起始机制和原核生物的翻译起始机制之间的惊人的相似之处进行了讨论。
Initiation of translation of hepatitis C virus and classical swine fever virus mRNAs results from internal ribosomal entry. We reconstituted internal ribosomal entry in vitro from purified translation components and monitored assembly of 48S ribosomal preinitiation complexes by toe-printing. Ribosomal subunits (40S) formed stable binary complexes on both mRNAs. The complex structure of these RNAs determined the correct positioning of the initiation codon in the ribosomal "P" site in binary complexes. Ribosomal binding and positioning on these mRNAs did not require the initiation factors eIF3, eIF4A, eIF4B, and eIF4F and translation of these mRNAs was not inhibited by a trans-dominant eIF4A mutant. Addition of Met-tRNA(i)(Met), eIF2, and GTP to these binary ribosomal complexes resulted in formation of 48S preinitiation complexes. The striking similarities between this eukaryotic initiation mechanism and the mechanism of translation initiation in prokaryotes are discussed.