Proteins surrounding hairpin IIIe of the hepatitis C virus internal ribosome entry site on the human 40S ribosomal subunit.
Proteins surrounding hairpin IIIe of the hepatitis C virus internal ribosome entry site on the human 40S ribosomal subunit.
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DOI：
10.1093/nar/gkl155
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发表时间：
2006
影响因子：
14.9
通讯作者：
Karpova G
中科院分区：
文献类型：
--
作者：
Laletina E;Graifer D;Malygin A;Ivanov A;Shatsky I;Karpova G
丙型肝炎病毒（HCV）RNA的内部核糖体进入位点（IRES）与无eIF的40 S核糖体亚基的结合是启动病毒RNA翻译的第一步。已知包含IRES的253-302 nt的发夹IIId和IIIe对于结合40 S亚基是必需的。在这里，我们研究了HCV IRES与人40 S核糖体亚基二元复合物的分子环境。为此目的，使用带有可光活化的全氟苯基叠氮化物交联剂的两种RNA衍生物。在一种衍生物中，交联剂位于发夹IIIe中的核苷酸A296处，而在另一种衍生物中，交联剂位于结构域II中的G87处。使用与靶RNA序列互补的寡脱氧核苷酸的烷基化衍生物进行交联剂的位点特异性引入。用任一RNA衍生物均未检测到与rRNA的交联。在A296处具有光活化基团的RNA与鉴定为S5和S16（主要）以及p40和S3 a（次要）的蛋白质交联，而在G87处修饰的RNA未检测到与蛋白质的交联。所获得的结果表明，发夹IIIe位于溶剂侧的40 S亚基头部的一个网站上的喙对面。
Binding of the internal ribosome entry site (IRES) of the hepatitis C virus (HCV) RNA to the eIF-free 40S ribosomal subunit is the first step of initiation of translation of the viral RNA. Hairpins IIId and IIIe comprising 253–302 nt of the IRES are known to be essential for binding to the 40S subunit. Here we have examined the molecular environment of the HCV IRES in its binary complex with the human 40S ribosomal subunit. For this purpose, two RNA derivatives were used that bore a photoactivatable perfluorophenyl azide cross-linker. In one derivative the cross-linker was at the nucleotide A296 in hairpin IIIe, and in the other at G87 in domain II. Site-specific introduction of the cross-linker was performed using alkylating derivatives of oligodeoxyribonucleotides complementary to the target RNA sequences. No cross-links with the rRNA were detected with either RNA derivative. The RNA with the photoactivatable group at A296 cross-linked to proteins identified as S5 and S16 (major) and p40 and S3a (minor), while no cross-links with proteins were detected with RNA modified at G87. The results obtained indicate that hairpin IIIe is located on the solvent side of the 40S subunit head on a site opposite the beak.
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DOI：
10.1016/s0167-4781(00)00252-9
发表时间：
2000-12-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION
影响因子：
--
作者：
Malygin, AA;Shaulo, DD;Karpova, GG
通讯作者：
Karpova, GG
DOI：
10.1111/j.1432-1033.1994.tb20100.x
发表时间：
1994-12-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
MALYGIN, AA;GRAIFER, DM;KARPOVA, GG
通讯作者：
KARPOVA, GG
影响因子：
64.5
作者：
Spahn, CMT;Beckmann, R;Frank, J
通讯作者：
Frank, J
影响因子：
5.6
作者：
Morgan, DG;Ménétret, JF;Akey, CW
通讯作者：
Akey, CW
影响因子：
4.8
作者：
Fukushi, S;Okada, M;Katayama, K
通讯作者：
Katayama, K