Attal J, Théron M C, Taboit F, Cajero-Juarez M, Kann G, Bolifraud P, Houdebine L M
Unité de Différenciation Cellulaire, Institut National de la Recherche Agronomique, Jouy-en-Josas, France.
FEBS Lett. 1996 Sep 2;392(3):220-4. doi: 10.1016/0014-5793(96)00815-0.
RNA fragments containing the complete R region and the beginning of the U5 region ('R') from the human T cell leukaemia virus 1 (HTLV-1) stimulated the translation of the second cistrons in bicistronic mRNAs. The 5' untranslated region from SV40 early genes (SU) which was unable to stimulate translation of second cistrons amplified markedly the internal ribosome entry site (IRES) effect of the HTLV-1 'R' fragments. The 'R' regions from HTLV-1 have therefore properties similar to internal ribosome entry sites (IRES) originally found in picornavirus. The beginning of the U5 region from HTLV-1 contains a polypyrimidine sequence which is known to play an essential role in the IRES activity in picornavirus. The same experiments carried out using the 'R' region from bovine leukaemia virus (BLV) showed that this sequence has at most a weak IRES effect. One retroviruses, HTLV-1 and perhaps others contain therefore an IRES activity. Interestingly, the combined SU 'R' sequence worked efficiently with different cistrons, different promoters and in all tested cell lines, whereas the poliovirus IRES was active in CHO cells but not in the mouse mammary cell line HC11. The SU 'R' sequence may therefore preferably be used to generate active bicistronic mRNAs.
含有来自人类T细胞白血病病毒1型(HTLV-1)完整R区域和U5区域起始部分(“R”)的RNA片段刺激了双顺反子mRNA中第二个顺反子的翻译。来自SV40早期基因的5'非翻译区(SU)虽然不能刺激第二个顺反子的翻译,但却显著增强了HTLV-1“R”片段的内部核糖体进入位点(IRES)效应。因此,HTLV-1的“R”区域具有与最初在小RNA病毒中发现的内部核糖体进入位点(IRES)相似的特性。HTLV-1的U5区域起始部分包含一个多嘧啶序列,已知该序列在小RNA病毒的IRES活性中起关键作用。使用牛白血病病毒(BLV)的“R”区域进行的相同实验表明,该序列至多具有微弱的IRES效应。因此,一种逆转录病毒,HTLV-1以及可能的其他病毒含有IRES活性。有趣的是,SU“R”组合序列能与不同的顺反子、不同的启动子有效配合,并且在所有测试的细胞系中均有效,而脊髓灰质炎病毒IRES在CHO细胞中具有活性,但在小鼠乳腺细胞系HC11中无活性。因此,SU“R”序列可能更适合用于产生有活性的双顺反子mRNA。