Leigh Jessica, Lang B Franz
Département de Biochimie, Université de Montréal, Montréal (Québec), H3T 1J4, Canada.
RNA. 2004 Apr;10(4):615-21. doi: 10.1261/rna.5195504.
The jakobid flagellates are bacteriovorus protists with mitochondrial genomes that are the most ancestral identified to date, in that they most resemble the genomes of the alpha-proteobacterial ancestors of the mitochondrion. Because of the bacterial character of jakobid mitochondrial genomes, it was expected that mechanisms for gene expression and RNA structures would be bacterial in nature. However, sequencing of the mitochondrial genome of the jakobid Seculamonas ecuadoriensis revealed several apparent mismatches in the acceptor stems of two predicted tRNAs. To investigate this observation, we determined the cDNA sequences of these tRNAs by RT-PCR. Our results show that the last three positions of the 3' extremity, plus the discriminator position of seryl and glutamyl tRNAs, are altered posttranscriptionally, restoring orthodox base-pairing and replacing the discriminator with an adenosine residue, in an editing process that resembles that of the metazoan Lithobius forficatus. However, the most 5' of the edited nucleotides is occasionally left unedited, indicating that the editing mechanism proceeds initially by exonucleolytic degradation, followed by repair of the degraded region. This 3' tRNA editing mechanism is likely distinct from that of L. forficatus, despite the apparent similarities between the two systems.
贾科比鞭毛虫是食细菌的原生生物,其线粒体基因组是迄今为止已鉴定出的最原始的,因为它们与线粒体的α-变形菌祖先的基因组最为相似。由于贾科比线粒体基因组具有细菌特征,人们预计其基因表达和RNA结构的机制本质上是细菌的。然而,对贾科比鞭毛虫厄瓜多尔单鞭滴虫线粒体基因组的测序揭示了两个预测的tRNA的接受茎中存在几个明显的错配。为了研究这一观察结果,我们通过RT-PCR确定了这些tRNA的cDNA序列。我们的结果表明,在类似于后生动物石蜈蚣的编辑过程中,3'末端的最后三个位置,加上丝氨酰和谷氨酰胺tRNA的鉴别位点,在转录后发生改变,恢复了正统的碱基配对,并用腺苷残基取代了鉴别位点。然而,编辑核苷酸中最靠近5'端的部分偶尔会未被编辑,这表明编辑机制最初是通过核酸外切酶降解进行的,随后是对降解区域的修复。尽管这两个系统之间有明显的相似之处,但这种3'tRNA编辑机制可能与石蜈蚣的不同。