Instituto de Ciencias Biomedicas, Facultad de Medicina y Facultad de Ciencias de la Vida, Universidad Andres Bello, Santiago 83700071, Chile.
Viruses. 2024 Mar 5;16(3):403. doi: 10.3390/v16030403.
LTR-retrotransposons are transposable elements characterized by the presence of long terminal repeats (LTRs) directly flanking an internal coding region. They share genome organization and replication strategies with retroviruses. Steamer-like Element-1 (SLE-1) is an LTR-retrotransposon identified in the genome of the Chilean blue mussel . SLE-1 is transcribed; however, whether its RNA is also translated and the mechanism underlying such translation remain to be elucidated. Here, we characterize the SLE-1 translation mechanism. We found that the SLE-1 5' and 3'LTRs command transcription of sense and antisense RNAs, respectively. Using luciferase reporters commanded by the untranslated regions (UTRs) of SLE-1, we found that in vitro 5'UTR sense is unable to initiate translation, whereas the antisense 5'UTR initiates translation even when the eIF4E-eIF4G interaction was disrupted, suggesting the presence of an internal ribosomal entry site (IRES). The antisense 5'UTR IRES activity was tested using bicistronic reporters. The antisense 5'UTR has IRES activity only when the mRNA is transcribed in the nucleus, suggesting that nuclear RNA-binding proteins are required to modulate its activity. Indeed, heterogeneous nuclear ribonucleoprotein K (hnRNPK) was identified as an IRES trans-acting factor (ITAF) of the SLE-1 IRES. To our knowledge, this is the first report describing an IRES in an antisense mRNA derived from a mussel LTR-retrotransposon.
长末端重复(LTR) retrotransposons 是一类可移动元件,其特征是在内部编码区的直接侧翼存在长末端重复(LTRs)。它们与逆转录病毒共享基因组组织和复制策略。Steamer-like Element-1(SLE-1)是在智利贻贝基因组中鉴定出的 LTR-retrotransposon。SLE-1 被转录;然而,其 RNA 是否也被翻译以及翻译的机制仍有待阐明。在这里,我们描述了 SLE-1 的翻译机制。我们发现,SLE-1 的 5'和 3'LTR 分别指挥 sense 和 antisense RNA 的转录。使用由 SLE-1 的非翻译区(UTR)命令的荧光素酶报告基因,我们发现体外 5'UTR sense 无法启动翻译,而 antisense 5'UTR 即使在 eIF4E-eIF4G 相互作用被破坏的情况下也启动翻译,表明存在内部核糖体进入位点(IRES)。使用双顺反子报告基因测试了 antisense 5'UTR 的 IRES 活性。只有当 mRNA 在核中转录时,antisense 5'UTR 才具有 IRES 活性,这表明需要核 RNA 结合蛋白来调节其活性。事实上,异质核核糖核蛋白 K(hnRNPK)被鉴定为 SLE-1 IRES 的 IRES 反式作用因子(ITAF)。据我们所知,这是首次描述来自贻贝 LTR-retrotransposon 的 antisense mRNA 中的 IRES。