Schmidt W M, Schweyen R J, Wolf K, Mueller M W
Vienna Biocenter, University of Vienna, Austria.
J Mol Biol. 1994 Oct 21;243(2):157-66. doi: 10.1006/jmbi.1994.1642.
The recent report on RNA-mediated group II intron (IVS, intervening sequence) transposition in mitochondria (mt) of Saccharomyces cerevisiae and Podospora anserina and the demonstration of reverse transcriptase (RT) activity encoded by the mobile S. cerevisiae intron cox1-aI1 suggests that group II introns constitute a new class of site-specific retro-like (retroid) elements. This is supported by the finding that the mitochondrial cob1-bI1 intron from the fission yeast Schizosaccharomyces pombe, encoding an RT-like open reading frame, is transposed in mtDNA populations. In agreement with the involvement of an RNA-intermediate in IVS transposition: First, the insertion sites were preceded by at least an IBS1-like (intron binding site) motif, which corresponds to the upstream exon and suffices to form the IBS1/EBS1 (EBS: exon binding site) base-pairing interactions. Second, intron transposition was conservative with respect to sequences flanking the insertion sites. We formulated the hypothesis that transient IVS insertion at non-allelic sites followed by recombination can be viewed as a general molecular mechanism, applicable equally well to site-specific genomic instabilities involving splice-site borders of group II introns and to the formation of extra-genomic IVS plasmid DNAs (plDNAs). We used polymerase chain reaction (PCR) techniques to detect infrequent rearrangements in mtDNA and report here on duplicative IVS transposition, twintron formation (e.g. bI1 insertion into another bI1 intron), and IVS insertions at canonical 5' exon-intron borders in S. pombe (cob1-bI1) and in S. cerevisiae (cox1-aI1). These data substantiate the concept that group II intron homing, IVS transposition and circular IVS plDNA formation involve a common RNA-mediated mechanism. Finally, the findings suggest that extra-genomic group II IVS copies are not restricted to senescence mycelia of P. anserina, but constitute natural components of group II IVS-containing genomes.
最近关于酿酒酵母和嗜热栖热放线菌线粒体(mt)中RNA介导的II类内含子(IVS,间隔序列)转座的报告,以及由可移动的酿酒酵母内含子cox1-aI1编码的逆转录酶(RT)活性的证明,表明II类内含子构成了一类新的位点特异性类逆转录(类逆转录)元件。这一观点得到了以下发现的支持:来自裂殖酵母粟酒裂殖酵母的线粒体cob1-bI1内含子编码一个类似RT的开放阅读框,它在mtDNA群体中发生转座。与RNA中间体参与IVS转座一致:首先,插入位点之前至少有一个类似IBS1(内含子结合位点)的基序,它对应于上游外显子,足以形成IBS1/EBS1(EBS:外显子结合位点)碱基配对相互作用。其次,内含子转座在插入位点侧翼序列方面是保守的。我们提出了一个假设,即非等位位点的瞬时IVS插入随后发生重组可以被视为一种普遍的分子机制,同样适用于涉及II类内含子剪接位点边界的位点特异性基因组不稳定性以及基因组外IVS质粒DNA(plDNA)的形成。我们使用聚合酶链反应(PCR)技术检测mtDNA中罕见的重排,并在此报告粟酒裂殖酵母(cob1-bI1)和酿酒酵母(cox1-aI1)中重复IVS转座、双内含子形成(例如bI1插入另一个bI1内含子)以及IVS在典型5'外显子-内含子边界处的插入。这些数据证实了II类内含子归巢、IVS转座和环状IVS plDNA形成涉及共同的RNA介导机制这一概念。最后,这些发现表明基因组外的II类IVS拷贝并不局限于嗜热栖热放线菌的衰老菌丝体,而是构成含有II类IVS基因组的天然成分。