Human hepatic glyceraldehyde-3-phosphate dehydrogenase binds to the poly(U) tract of the 3′ non-coding region of hepatitis C virus genomic RNA
Human hepatic glyceraldehyde-3-phosphate dehydrogenase binds to the poly(U) tract of the 3′ non-coding region of hepatitis C virus genomic RNA
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DOI：
10.1099/0022-1317-80-12-3109
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发表时间：
1999-12-01
影响因子：
3.8
通讯作者：
Alexander, GJM
中科院分区：
文献类型：
--
作者：
Petrik, J;Parker, H;Alexander, GJM
丙型肝炎病毒（HCV）基因组RNA的3'端非编码区（3' NCR）的中间部分是一个独特的poly（U/UC）区，它可能是HCV复制酶复合物的识别信号。在这项研究中，使用HCV阴性或阳性肝外植体的胞质提取物鉴定了几种与模拟这种结构的固定化核糖寡核苷酸r（U）（25）特异性结合的蛋白质，并进一步研究了一种突出的36 kDa蛋白质。包括均聚物的竞争实验揭示了结合亲和力的顺序：oligo/poly（U）>（A）>（C）>（G）。该蛋白被鉴定为甘油醛-3-磷酸脱氢酶（GAPDH），已知其是结合RNA的多功能蛋白。GAPDH有效地结合全长HCV RNA和结合各种3 'NCR构建体揭示了对3' NCR的中间部分的存在的关键依赖性，先前描述的结合3 'NCR的多嘧啶片段结合蛋白不能有效地结合3' NCR的中间部分，并且以相当小的量从肝提取物中捕获。
The unique poly(U/UC) tract, the middle part of the tripartite 3' non-coding region (3'NCR) of hepatitis C virus (HCV) genomic RNA, may represent a recognition signal for the HCV replicase complex. In this study, several proteins binding specifically to immobilized ribooligonucleotide r(U)(25) mimicking this structure were identified using cytosolic extracts from HCV-negative or -positive liver explants, and a prominent 36 kDa protein was studied further. Competition experiments including homoribopolymers revealed binding affinities in the order: oligo/poly(U)>(A)>(C)>(G), The protein was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a multifunctional protein known to bind RNA. GAPDH bound efficiently to the full-length HCV RNA and binding to various 3'NCR constructs revealed critical dependence upon the presence of the middle part of the 3'NCR, Polypyrimidine tract-binding protein, described previously to bind the 3'NCR, did not bind efficiently to the middle part of 3'NCR and was captured from liver extracts in considerably smaller quantities.