Munroe S H
Nucleic Acids Res. 1984 Nov 26;12(22):8437-56. doi: 10.1093/nar/12.22.8437.
In order to investigate the possible role of RNA secondary structure in determining the efficiency and specificity of mRNA splicing, the structures of sequences at three acceptor splice sites in adenovirus were studied. Transcripts spanning intron-exon junctions were synthesized using SP6 RNA polymerase and analyzed using single and double-strand specific nucleases. Distinctive patterns of nuclease cleavage were observed for each of the 3 sites examined. At both sites in the E2a region sequences adjacent to the splice sites were particularly susceptible to digestion with T1 and S1 nucleases. In contrast, a splice site for hexon mRNA was largely resistant to these nucleases. The results obtained suggest that the conformation of the RNA at some, but not all, acceptor sites may enhance the accessibility of these sites to factors involved in splicing nuclear RNA and confirm the presence of a large, previously predicted hairpin structure centered on the acceptor site at 67 map units.
为了研究RNA二级结构在决定mRNA剪接效率和特异性方面可能发挥的作用，对腺病毒中三个受体剪接位点处序列的结构进行了研究。使用SP6 RNA聚合酶合成跨越内含子-外显子连接的转录本，并使用单链和双链特异性核酸酶进行分析。在所检测的3个位点中，每个位点都观察到了独特的核酸酶切割模式。在E2a区域的两个位点，与剪接位点相邻的序列特别容易被T1和S1核酸酶消化。相比之下，六邻体mRNA的一个剪接位点对这些核酸酶具有很大的抗性。所得结果表明，在一些但并非所有受体位点处，RNA的构象可能会增强这些位点对参与核RNA剪接的因子的可及性，并证实存在一个以67个图谱单位处的受体位点为中心的、先前预测的大的发夹结构。
