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首页> 外文期刊>Molecular and Cellular Biology >Sequences homologous to 5' splice sites are required for the inhibitory activity of papillomavirus late 3' untranslated regions.
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Sequences homologous to 5' splice sites are required for the inhibitory activity of papillomavirus late 3' untranslated regions.

机译：乳头瘤病毒晚期3'非翻译区的抑制活性需要与5'剪接位点同源的序列。

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Expression of bovine papillomavirus type 1 (BPV-1) late genes is limited to terminally differentiated keratinocytes in an infected epithelium. We have previously shown that although the BPV-1 late polyadenylation site is functional in nonpermissive cells, a 53-nucleotide (nt) fragment of the late 3' untranslated region acts posttranscriptionally to reduce polyadenylated cytoplasmic RNA levels. This 53-nt fragment does not appear to function by destabilizing polyadenylated cytoplasmic RNA (P. A. Furth and C. C. Baker, J. Virol. 65:5806-5812, 1991). In this study, we used site-directed mutagenesis and deletion analysis to demonstrate that the sequence AAG/GUAAGU, which is identical to the consensus 5' splice site sequence, was both necessary and sufficient for the inhibitory activity of the 53-nt fragment. Furthermore, base pairing between the 5' end of the U1 small nuclear RNA and this 5' splice site-like sequence was shown to be required for the inhibitory activity in vivo. We have also further mapped the human papillomavirus type 16 late 3' inhibitory element (I. M. Kennedy, J. K. Haddow, and J. B. Clements, J. Virol. 65:2093-2097, 1991) to a 51-nt region containing four overlapping sequence motifs with partial homology to 5' splice sites. Mutation of each of these motifs demonstrated that only one of these motifs is required for the inhibitory activity. However, the presence of the other motifs may contribute to the full inhibitory activity of the element. No BPV-1 or human papillomavirus type 16 mRNAs which are spliced by using the potential 5' splice sites present in the viral late 3' untranslated regions have been identified. This suggests that the primary function of these 5' splice site-like sequences is the inhibition of late gene expression. The most likely mechanism of action of these elements is reduction of polyadenylation efficiency, perhaps through interference with 3'-terminal exon definition.
机译：牛乳头瘤病毒1型（BPV-1）晚期基因的表达仅限于感染上皮细胞中的终末分化角质形成细胞。我们先前已经表明，尽管BPV-1晚期聚腺苷酸化位点在非允许细胞中具有功能，但晚期3'非翻译区的53个核苷酸（nt）片段在转录后起作用，以降低聚腺苷酸化的细胞质RNA水平。该53-nt片段似乎没有通过使聚腺苷酸化的细胞质RNA去稳定而起作用（P.A.Furth和C.C.Baker，J.Virol.65：5806-5812，1991）。在这项研究中，我们使用了定点诱变和缺失分析，以证明序列AAG / GUAAGU与共有的5'剪接位点序列相同，对于53 nt片段的抑制活性既必要又充分。此外，显示U1小核RNA的5'末端与该5'剪接位点样序列之间的碱基配对是体内抑制活性所必需的。我们还将人乳头瘤病毒16型晚期3'抑制元件（IM Kennedy，JK Haddow，and JB Clements，J.Virol。65：2093-2097，1991）定位到一个51 nt区域，该区域包含四个具有重叠序列的基序，与5'剪接位点部分同源。这些基序中的每一个的突变表明抑制活性仅需要这些基序中的一个。但是，其他基序的存在可能有助于元素的完全抑制活性。尚未发现通过使用病毒晚期3'非翻译区中潜在的5'剪接位点剪接的BPV-1或人类乳头瘤病毒16型mRNA。这表明这些5'剪接位点样序列的主要功能是抑制晚期基因表达。这些元素最可能的作用机制可能是通过干扰3'-末端外显子的定义来降低聚腺苷酸化效率。

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