A CONSERVED HELICAL ELEMENT IS ESSENTIAL FOR INTERNAL INITIATION OF TRANSLATION OF HEPATITIS-C VIRUS-RNA

被引:136
作者
WANG, CY
SARNOW, P
SIDDIQUI, A
机构
[1] UNIV COLORADO,SCH MED,DEPT MICROBIOL,DENVER,CO 80262
[2] UNIV COLORADO,SCH MED,DEPT BIOCHEM BIOPHYS & GENET,DENVER,CO 80262
[3] UNIV COLORADO,SCH MED,PROGRAM MOLEC BIOL,DENVER,CO 80262
关键词
D O I
10.1128/JVI.68.11.7301-7307.1994
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Translation of hepatitis C virus (HCV) RNA is initiated by cap-independent internal ribosome binding to the 5' noncoding region (NCR). To identify the sequences and structural elements within the 5' NCR of HCV RNA that contribute to the initiation of translation, a series of point mutations was introduced within this sequence. Since the pyrimidine-rich tract is considered a characteristic feature of picornavirus internal ribosome entry site (IRES) elements, our mutational analysis focused on two putative pyrimidine tracts (Py-I and Py-II) within the HCV 5' NCR, Translational efficiency of these mutant RNAs was examined by in vitro translation and after RNA transfection into liver-derived cells. Mutational analysis of Py-I (nucleotides 120 to 130), supported by compensatory mutants, demonstrates that the primary sequence of this motif is not important but that a helical structural element associated with this region is critical for HCV IRES function. Mutations in Py-II (nucleotides 191 to 199) show that this motif is dispensable for IRES function as well. Thus, the pyrimidine-rich tract motif, which is considered as an essential element of the picornavirus IRES elements, does not appear to be a functional component of the HCV IRES. Further, the insertional mutagenesis study suggests a requirement for proper spacing between the initiator AUG and the upstream structures of the HCV IRES element for internal initiation of translation.
引用
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页码:7301 / 7307
页数:7
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