School of Biology, University of St. Andrews, Biomolecular Sciences Research Complex, North Haugh, St. Andrews KY16 9ST, UK.
International College, National Pingtung University of Science and Technology, 1, Shuefu Rd., Neipu, Pingtung 91201, Taiwan.
Viruses. 2024 Oct 9;16(10):1587. doi: 10.3390/v16101587.
Alignment of picornavirus proteinase/polymerase sequences reveals this family evolved into five 'supergroups'. Interestingly, the nature of the 2A region of the picornavirus polyprotein is highly correlated with this phylogeny. Viruses within supergroup 4, the , have complex 2A regions with many viruses encoding multiple 2A sequences. In vitro transcription/translation analyses of a synthetic polyprotein comprising green fluorescent protein (GFP) linked to β-glucuronidase (GUS) via individual 2As showed two main phenotypes: highly active 2A sequences-similar to foot-and-mouth disease virus 2A-and, surprisingly, a novel phenotype of some 2A sequences which apparently terminate translation at the C-terminus of 2A without detectable re-initiation of downstream sequences (GUS). Probing databases with the short sequences between 2As did not reveal any potential 'accessory' functions. The novel, highly active, 2A-like sequences we identified substantially expand the toolbox for biomedical/biotechnological co-expression applications.
小核糖核酸病毒蛋白酶/聚合酶序列的排列揭示了这个家族进化成了五个“超级组”。有趣的是，小核糖核酸病毒多蛋白的 2A 区的性质与这个系统发育高度相关。超级组 4 中的病毒具有复杂的 2A 区，许多病毒编码多个 2A 序列。通过单个 2A 将绿色荧光蛋白（GFP）与β-葡糖苷酸酶（GUS）连接的合成多蛋白的体外转录/翻译分析显示出两种主要表型：高度活跃的 2A 序列-类似于口蹄疫病毒 2A-，令人惊讶的是，一些 2A 序列表现出新颖的表型，它们显然在 2A 的 C 末端终止翻译，而下游序列（GUS）没有可检测到的重新起始。用 2A 之间的短序列探测数据库并没有揭示任何潜在的“辅助”功能。我们鉴定的新型、高度活跃的 2A 样序列大大扩展了用于生物医学/生物技术共表达应用的工具包。
