Wakita T, Wands J R
Molecular Hepatology Laboratory, Massachusetts General Hospital Cancer Center, Boston.
J Biol Chem. 1994 May 13;269(19):14205-10.
The effect of sense and antisense oligodeoxynucleotides (ODNs) on hepatitis C virus (HCV) gene expression was studied to determine the role of the highly conserved 5'-untranslated region in the life cycle of the virus. It was found that antisense ODNs complementary to nucleotides (nt) 38-65, 134-175, and 312-339 in the 5' noncoding region and 341-377 in the core open reading frame efficiently blocked HCV RNA translation. Overlapping ODNs that differed by only several nucleotides showed substantially different inhibition of HCV RNA translation. Fine sequence specificity testing at nt positions 351-377 revealed that ODNs as small as a 12-mer (nt 351-363) retained a high degree (80%) of inhibitory activity compared to ODNs of longer sequences. These results suggest that there are three highly specific domains in the 5' noncoding region and a sequence immediately downstream of the HCV core initiation codon that may be critical for translation of HCV RNA. This study also provides an experimental approach for the selection of target HCV RNA sequences susceptible to antisense effects, as well as for definition of functional regions of the genome necessary for viral replication.
研究了正义和反义寡脱氧核苷酸（ODNs）对丙型肝炎病毒（HCV）基因表达的影响，以确定高度保守的5'非翻译区在病毒生命周期中的作用。发现与5'非编码区的核苷酸（nt）38 - 65、134 - 175和312 - 339以及核心开放阅读框中的341 - 377互补的反义ODNs能有效阻断HCV RNA翻译。仅相差几个核苷酸的重叠ODNs对HCV RNA翻译的抑制作用有很大差异。在nt位置351 - 377进行的精细序列特异性测试表明，与较长序列的ODNs相比，小至12聚体（nt 351 - 363）的ODNs仍保留高度（80%）的抑制活性。这些结果表明，在5'非编码区有三个高度特异性的结构域以及HCV核心起始密码子下游紧邻的一个序列，它们可能对HCV RNA的翻译至关重要。本研究还为选择易受反义效应影响的靶HCV RNA序列以及确定病毒复制所需的基因组功能区域提供了一种实验方法。
