Department of Laboratory Medicine, Lund University, BMC-B13, 223 62 Lund, Sweden.
Department of Laboratory Medicine, Lund University, BMC-B13, 223 62 Lund, Sweden.
Virus Res. 2018 Jan 15;244:128-136. doi: 10.1016/j.virusres.2017.11.009. Epub 2017 Nov 16.
The HPV16 E5 open reading frame (ORF) is present on the majority of all alternatively spliced HPV16 mRNAs, but it is currently unknown how well it is translated into E5 protein. To identify HPV16 mRNAs that are efficiently translated into E5, we have generated cDNA plasmids expressing individual, alternatively spliced HPV16 mRNAs with the potential to produce E5. By replacing the E5 ORF with sLuc, we could quantitate sLuc and determine how well each cDNA was translated. Our results showed that the upstream E1 and E7 AUGs inhibited translation of the E5 ORF and revealed that only one HPV16 mRNA produced high levels of E5. This was an HPV16 early mRNA spliced from SD226 to SA3358. These results were confirmed in the context of the entire HPV16 genome. Taken together, our results indicate that E5 is expressed early in the HPV16 replication cycle since it is translated efficiently only by one early mRNA.
HPV16E5 开放阅读框（ORF）存在于大多数替代拼接的 HPV16mRNA 中，但目前尚不清楚其翻译成 E5 蛋白的效率如何。为了鉴定能够有效翻译成 E5 的 HPV16mRNA，我们生成了表达具有产生 E5 潜力的单个替代拼接 HPV16mRNA 的 cDNA 质粒。通过用 sLuc 替换 E5ORF，我们可以定量 sLuc，并确定每个 cDNA 的翻译效率。我们的结果表明，上游 E1 和 E7AUG 抑制了 E5ORF 的翻译，并揭示只有一种 HPV16mRNA 产生高水平的 E5。这是一种从 SD226 拼接至 SA3358 的 HPV16 早期 mRNA。在整个 HPV16 基因组的背景下，这些结果得到了证实。总之，我们的结果表明，E5 在 HPV16 复制周期的早期表达，因为它仅由一种早期 mRNA 有效翻译。
