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帽结合蛋白复合物真核起始因子(eIF)4F和eIF(iso)4F与烟草坏死卫星病毒3'非翻译区一个区域的新型相互作用。

A novel interaction of Cap-binding protein complexes eukaryotic initiation factor (eIF) 4F and eIF(iso)4F with a region in the 3'-untranslated region of satellite tobacco necrosis virus.

作者信息

Gazo Brandy M, Murphy Patricia, Gatchel Jennifer R, Browning Karen S

机构信息

Department of Chemistry and Biochemistry and the Institute for Cellular and Molecular Biology, University of Texas, Austin, Texas 78712, USA.

出版信息

J Biol Chem. 2004 Apr 2;279(14):13584-92. doi: 10.1074/jbc.M311361200. Epub 2004 Jan 17.

DOI:10.1074/jbc.M311361200
PMID:14729906
Abstract

Satellite tobacco necrosis virus (STNV) RNA is naturally uncapped at its 5' end and lacks polyadenylation at its 3' end. Despite lacking these two hallmarks of eukaryotic mRNAs, STNV-1 RNA is translated very efficiently. A approximately 130-nucleotide translational enhancer (TED), located 3' to the termination codon, is necessary for efficient cap-independent translation of STNV-1 RNA. The STNV-1 TED RNA fragment binds to the eukaryotic cap-binding complexes, initiation factor (eIF) 4F and eIF(iso)4F, as measured by nitrocellulose binding and fluorescence titration. STNV-1 TED is a potent inhibitor of in vitro translation when added in trans. This inhibition is reversed by the addition of eIF4F or eIF(iso)4F, and the subunits of eIF4F and eIF(iso)4F cross-link to STNV-1 TED, providing additional evidence that these factors interact directly with STNV-1 TED. Deletion mutagenesis of the STNV-1 TED indicates that a minimal region of approximately 100 nucleotides is necessary to promote cap-independent translation primarily through interaction with the cap binding subunits (eIF4E or eIF(iso)4E) of eIF4F or eIF(iso)4F.

摘要

卫星烟草坏死病毒(STNV)RNA在其5'端天然无帽,在其3'端缺乏多聚腺苷酸化。尽管缺乏真核mRNA的这两个特征,但STNV-1 RNA的翻译效率非常高。位于终止密码子下游3'端的一个约130个核苷酸的翻译增强子(TED),是STNV-1 RNA高效的不依赖帽的翻译所必需的。通过硝酸纤维素结合和荧光滴定测量,STNV-1 TED RNA片段与真核帽结合复合物、起始因子(eIF)4F和eIF(iso)4F结合。当反式添加时,STNV-1 TED是体外翻译的有效抑制剂。添加eIF4F或eIF(iso)4F可逆转这种抑制作用,并且eIF4F和eIF(iso)4F的亚基与STNV-1 TED交联,提供了这些因子与STNV-1 TED直接相互作用的额外证据。STNV-1 TED的缺失诱变表明,一个约100个核苷酸的最小区域主要通过与eIF4F或eIF(iso)4F的帽结合亚基(eIF4E或eIF(iso)4E)相互作用来促进不依赖帽的翻译是必要的。

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