Bhat B M, Brady H A, Pursley M H, Wold W S
J Mol Biol. 1986 Aug 20;190(4):543-57. doi: 10.1016/0022-2836(86)90240-8.
Region E3 of the adenovirus encodes about ten overlapping mRNAs (a to j) with different splicing patterns and with two RNA 3' end sites termed E3A and E3B. We have examined how deletions in 12 viable virus mutants affect differential RNA processing in E3. We assayed E3 mRNAs by the nuclease-gel and RNA blot procedures. Some deletions had no effect whereas others (e.g. deletion of a 3' splice or the E3A 3' end signal) had the anticipated effects on RNA processing. However, deletions in two regions had surprising effects. Deletions in one region (nucleotides 1691 to 2044) enhanced splicing at the upstream 951 5' splice site and the downstream 2157 and/or 2880 3' splice sites. Some of these deletions prevented RNA 3' end formation at the downstream E3A site. Deletion in the other region (nucleotides 2173 to 2237) enhanced an upstream splice site (951 to 2157) such that almost all pre-mRNA was processed into mRNA f. We suggest that these two regions contain cis-acting signals that regulate differential RNA processing. We discuss the results in terms of RNA folding and scanning models for splicing, as well as models for differential RNA 3' end formation at the E3A versus the E3B site.
腺病毒的E3区域编码约十种具有不同剪接模式的重叠mRNA(a至j),并具有两个称为E3A和E3B的RNA 3'末端位点。我们研究了12个存活病毒突变体中的缺失如何影响E3中的差异RNA加工。我们通过核酸酶凝胶和RNA印迹程序检测E3 mRNA。一些缺失没有影响,而其他缺失(例如3'剪接或E3A 3'末端信号的缺失)对RNA加工有预期的影响。然而,两个区域的缺失产生了惊人的影响。一个区域(核苷酸1691至2044)的缺失增强了上游951 5'剪接位点以及下游2157和/或2880 3'剪接位点的剪接。其中一些缺失阻止了下游E3A位点的RNA 3'末端形成。另一个区域(核苷酸2173至2237)的缺失增强了上游剪接位点(951至2157),使得几乎所有前体mRNA都加工成了mRNA f。我们认为这两个区域包含调节差异RNA加工的顺式作用信号。我们根据剪接的RNA折叠和扫描模型以及E3A与E3B位点处差异RNA 3'末端形成的模型来讨论这些结果。