Cermakian N, Ikeda T M, Cedergren R, Gray M W
Canadian Institute for Advanced Research, Department de Biochimie, Universite de Montreal, Quebec, Canada.
Nucleic Acids Res. 1996 Feb 15;24(4):648-54. doi: 10.1093/nar/24.4.648.
Although mitochondria and chloroplasts are considered to be descendants of eubacteria-like endo- symbionts, the mitochondrial RNA polymerase of yeast is a nucleus-encoded, single-subunit enzyme homologous to bacteriophage T3 and T7 RNA polymerases, rather than a multi-component, eubacterial-type alpha 2 beta beta' enzyme, as encoded in chloroplast DNA. To broaden our knowledge of the mitochondrial transcriptional apparatus, we have used a polymerase chain reaction (PCR) approach designed to amplify an internal portion of phage T3/T7-like RNA polymerase genes. Using this strategy, we have recovered sequences homologous to yeast mitochondrial and phage T3/T7 RNA polymerases from a phylogenetically broad range of multicellular and unicellular eukaryotes. These organisms display diverse patterns of mitochondrial genome organization and expression, and include species that separated from the main eukaryotic line early in the evolution of this lineage. In certain cases, we can deduce that PCR-amplified sequences, some of which contain small introns, are localized in nuclear DNA. We infer that the T3/T7-like RNA polymerase sequences reported here are likely derived from genes encoding the mitochondrial RNA polymerase in the organisms in which they occur, suggesting a phage T3/T7-like RNA polymerase was recruited to act in transcription in the mitochondrion at an early stage in the evolution of this organelle.
尽管线粒体和叶绿体被认为是类似真细菌的内共生体的后代,但酵母的线粒体RNA聚合酶是一种由细胞核编码的单亚基酶,与噬菌体T3和T7 RNA聚合酶同源,而不是像叶绿体DNA中编码的那样是一种多组分的真细菌型α2ββ'酶。为了拓宽我们对线粒体转录装置的认识,我们采用了一种聚合酶链反应(PCR)方法,旨在扩增噬菌体T3/T7样RNA聚合酶基因的内部片段。利用这一策略,我们从系统发育范围广泛的多细胞和单细胞真核生物中获得了与酵母线粒体和噬菌体T3/T7 RNA聚合酶同源的序列。这些生物表现出线粒体基因组组织和表达的多样模式,包括在真核生物谱系进化早期就与主要真核生物谱系分离的物种。在某些情况下,我们可以推断PCR扩增的序列,其中一些含有小内含子,定位于核DNA中。我们推断,这里报道的T3/T7样RNA聚合酶序列可能源自其所在生物中编码线粒体RNA聚合酶的基因,这表明在该细胞器进化的早期阶段,一种噬菌体T3/T7样RNA聚合酶被招募来参与线粒体的转录。