Winter A J, van der Horst G, Tabak H F
Laboratory of Biochemistry, University of Amsterdam, The Netherlands.
Nucleic Acids Res. 1988 May 11;16(9):3845-61. doi: 10.1093/nar/16.9.3845.
We have characterized the in vitro self-splicing of intron aI5 alpha containing precursor RNA from the yeast mitochondrial gene coding for cytochrome oxidase subunit I. This intron follows the rules for group I self-splicing introns and all the characteristic products have been identified. In addition we have detected abnormal RNA products with features that indicate that the self-splicing behaviour of this intron is more complex. Two intron circles are formed by use of a major and minor intron-internal site for circle closure. A cryptic 5'-splice site located in the 3' exon results in guanosine nucleotide mediated opening at a position 30 nt downstream of the normal 3' splice site. The reactions can all be explained on the basis of the "splice guide" model proposed by Davies et al (1982 Nature 300 719-724). Although the sequence motifs at cyclization and splice sites occur more often in this intron, only some of them are allowed to interact with the internal guide sequence, suggesting that both primary structure and spatial folding of the RNA are involved in formation of productive reaction sites.
我们已经对来自酵母线粒体基因中编码细胞色素氧化酶亚基I的含前体RNA的内含子aI5α的体外自我剪接进行了表征。该内含子遵循I类自我剪接内含子的规则,并且所有特征性产物均已得到鉴定。此外,我们还检测到了具有表明该内含子自我剪接行为更为复杂特征的异常RNA产物。通过使用主要和次要的内含子内部位点进行环化,形成了两个内含子环。位于3'外显子中的一个隐蔽的5'剪接位点导致鸟苷核苷酸在正常3'剪接位点下游30个核苷酸处介导的开放。所有这些反应都可以基于戴维斯等人(1982年,《自然》300 719 - 724)提出的“剪接引导”模型来解释。尽管环化和剪接位点处的序列基序在该内含子中出现得更为频繁,但只有其中一些能够与内部引导序列相互作用,这表明RNA的一级结构和空间折叠都参与了有效反应位点的形成。