ENTEROVIRUS 71 LEADER: TARGET FOR PEPTIDE INHIBITORS

每年都有成千上万的儿童感染肠病毒71型（EV71）。感染EV71病毒可导致严重并发症，如脊髓灰质炎样瘫痪、脑炎、脑膜炎甚至死亡。目前，没有针对EV71的治疗方法或疫苗。EV71属于小核糖核酸病毒家族，其成员含有正链RNA基因组，通过一种不寻常的内部核糖体进入机制进行翻译。研究人员将利用大多数细胞mRNA和病毒mRNA的翻译起始模型之间的差异来鉴定能够选择性抑制EV71 mRNA翻译的小肽。具体来说，表达构象受限肽库的逆转录病毒将被用于感染表达与增强型绿色荧光蛋白（EGFP）报告基因相关的ev715 ‘非编码区和与增强型黄色荧光蛋白（EYFP）相关的细胞c-myc 5’非编码区的细胞。通过细胞分选分离出不能表达EGFP但仍能表达EYFP的感染细胞，并分离编码推定抑制剂的基因。研究抑制肽的靶标和表征它们所破坏的分子相互作用将揭示更多关于病毒如何招募宿主细胞分子进行翻译的信息。抑制病毒RNA基因组扩增的细胞内稳定肽的选择和表征应该是寻找抗病毒治疗方法的新途径。

DESCRIPTION Every year hundreds of thousands of children are infected with enterovirus 71 (EV71). Infection with EV71 can lead to serious complications such as polio-like paralysis, encephalitis, meningitis or even death. Currently, there is no treatment or vaccine against EV71. EV71 belongs to the picornavirus family, whose members contain positive-stranded RNA genomes that are translated by an unusual mechanism of internal ribosome entry. The differences between the model of translational initiation of most cellular and viral mRNAs will be exploited to identify small peptides that can selectively inhibit the translation of the EV71 mRNA. Specifically, retroviruses expressing conformationally constrained peptide libraries will be used to infect cells expressing the EV71 5' non-coding region linked to an enhanced green fluorescent protein (EGFP) reporter gene and a cellular c-myc 5' non- coding region linked to an enhanced yellow fluorescence protein (EYFP). Infected cells which fail to express EGFP but still express EYFP will be isolated by cell sorting, and the gene encoding the putative inhibitor will be isolated . Studying the targets of the inhibitory peptides and characterizing the molecular interactions that they are disrupting will reveal more about how viruses function to recruit host cell molecules for their translation. Selection and characterization of intracellularly stable peptides that can inhibit the amplification of a viral RNA genome should level to novel ways in the search for antiviral therapeutics.