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从交联数据揭示的翻译中的人核糖体解码位点处围绕mRNA的18S rRNA核苷酸。

Nucleotides of 18S rRNA surrounding mRNA at the decoding site of translating human ribosome as revealed from the cross-linking data.

作者信息

Matassova N B, Venjaminova A G, Karpova G G

机构信息

Laboratory of Ribosomal Structure and Functions, Novosibirsk Institute of Bioorganic Chemistry, Siberian Branch of the Russian Academy of Sciences, Prospekt Lavrentieva, 8, 630090, Novosibirsk, Russian Federation.

出版信息

Biochim Biophys Acta. 1998 Apr 29;1397(2):231-9. doi: 10.1016/s0167-4781(98)00015-3.

Abstract

mRNA analogs, 4-(N-2-chloroethyl-N-methylamino)benzylmethyl-[5'-32P]-phosphamide derivatives of oligoribonucleotides pAUGUn (n=0, 3 or 6), were used for affinity labelling of human 80S ribosomes in complexes with codon-anticodon interaction at the P-site. These complexes were obtained in the presence of fractionated lysate from rabbit reticulocytes deprived of endogenous ribosomes and mRNAs. In all cases, 40S subunits were labelled preferentially. Within the subunits, both ribosomal proteins and 18S rRNA were modified. Ribosomal proteins cross-linked to pAUGUn derivatives were identified earlier. In this paper, nucleotides G-1010, G-1029, G-1033, G-1051, G-1054 and G-1059 of 18S rRNA cross-linked to both pAUG and pAUGU3 derivatives were identified by reverse transcription analysis.

摘要

mRNA类似物,即寡核糖核苷酸pAUGUn(n = 0、3或6)的4-(N-2-氯乙基-N-甲基氨基)苄基甲基-[5'-32P]-磷酰胺衍生物,用于在P位点与密码子-反密码子相互作用的复合物中对人80S核糖体进行亲和标记。这些复合物是在缺乏内源性核糖体和mRNA的兔网织红细胞分级裂解物存在下获得的。在所有情况下,40S亚基都被优先标记。在亚基内部,核糖体蛋白和18S rRNA都被修饰。之前已鉴定出与pAUGUn衍生物交联的核糖体蛋白。在本文中,通过逆转录分析鉴定了与pAUG和pAUGU3衍生物都交联的18S rRNA的核苷酸G-1010、G-1029、G-1033、G-1051、G-1054和G-1059。

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