The effects of nucleotide usage in key nucleotide positions+4 and-3 flanking start codon on translation levels mediated by IRES of hepatitis C virus
文献类型: 外文期刊
第一作者: Ma, P.
作者: Ma, P.;Ma, X-X;Chang, Q-Y;Li, L-J;Feng, Y-P;Ma, Z-R;Wang, Y-N;Zhou, J-H
作者机构:
关键词: Cap-independent translation; internal ribosomal entry site; hepatitis C virus; bicistronic expression vector; translation efficiency
期刊名称:ACTA VIROLOGICA ( 影响因子:1.162; 五年影响因子:1.255 )
ISSN: 0001-723X
年卷期: 2018 年 62 卷 4 期
页码:
收录情况: SCI
摘要: Internal ribosomal entry site (IRES) functions as a cis-acting RNA element, which drives an alternative and cap-independent translation initiation pathway. Currently, there are few studies on effects of nucleotide usages at key nucleotide positions +4 and -3 flanking start codon mediated by IRES of hepatitis C virus (HCV). Herein, we focus on the effect of nucleotide usages at -3 and +4 positions mediated by HCV IRES. The nucleotide contexts flanking AUG start codon employed by HCV IRES is firstly analyzed. We found that each position in the six nucleotide positions (-4 to +6) flanking start codon of HCV has a strong tendency to select the specific nucleotide. A set of bicistronic expression vectors containing CAT gene, HCV IRES and EGFP gene were constructed, including 16 different nucleotide combinations at position -3 and +4. Each set, in which nucleotide at the -3 and +4 position has been changed into different nucleotides, included 16 types of bicistronic expression vectors. It was found that the purine nucleotide at the position -3 or +4 obviously impacts on HCV IRES-related expression, and IRES-driven translation is potentially influenced by the Kozak rule. Our results suggest that optimization of nucleotides at positions -3 and +4 is a convenient and efficient way to enhance the level of IRES-mediated translation.
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