Children's Memorial Research Center, Immunology Department, 2300 Children's Plaza, Mailstop 212, Chicago, Illinois 60614, USA.
Mycologia. 2010 Nov-Dec;102(6):1288-94. doi: 10.3852/10-019. Epub 2010 Apr 26.
An open reading frame (ORF) was found in the mitochondrial genome of the Pan2-16 strain of Didymium iridis that showed high similarity to the NADH dehydrogenase subunit 3 (nad3) gene in other organisms. So far all other typical mitochondrial genes identified in this organism require RNA editing to generate ORFs capable of directing protein synthesis. The D. iridis sequence was compared to the putative nad3 gene in the related myxomycete Physarum polycephalum, which would require editing. Based on this comparison, editing sites could be predicted for the P. polycelphalum gene that would result in the synthesis of a highly conserved ND3 protein between the two organisms. To determine the editing status of the nad3 gene in other D. iridis strains, PCR was used to amplify this region from eight other independent isolates of the A1 Central American interbreeding series. In each case a 378 base pair ORF was detected by PCR amplification and sequencing. Three patterns of sequence variation were observed; however all base substitutions were in the third codon position and silent with respect to the amino acids encoded. The distribution of the sequence variants was mapped geographically. The requirement for RNA editing in all other typical mitochondrial genes of D. iridis and P. polycephalum and the presence of RNA editing in the nad3 gene of P. polycephalum suggest that the D. iridis nad3 gene might have been edited at one time. We propose that the D. iridis nad3 gene may have lost the requirement for RNA editing by reverse transcription of an edited transcript that subsequently was inserted into the genome.
在 Pan2-16 菌株的虹膜双腔菌的线粒体基因组中发现了一个开放阅读框(ORF),它与其他生物体中的 NADH 脱氢酶亚单位 3(nad3)基因具有高度相似性。到目前为止,在该生物体中鉴定的所有其他典型线粒体基因都需要 RNA 编辑才能产生能够指导蛋白质合成的 ORF。将 D. iridis 序列与相关粘菌多形肾形虫的假定 nad3 基因进行比较,该基因需要编辑。基于这种比较,可以预测 P. polycelphalum 基因的编辑位点,这将导致两个生物体之间合成高度保守的 ND3 蛋白。为了确定其他 D. iridis 菌株中 nad3 基因的编辑状态,使用 PCR 从 A1 中美洲杂交系列的另外 8 个独立分离株中扩增该区域。在每种情况下,通过 PCR 扩增和测序检测到 378 个碱基对的 ORF。观察到三种序列变异模式;然而,所有碱基替换都位于第三个密码子位置,并且相对于编码的氨基酸是沉默的。序列变异的分布进行了地理定位。D. iridis 和 P. polycephalum 的所有其他典型线粒体基因都需要 RNA 编辑,并且 P. polycephalum 的 nad3 基因存在 RNA 编辑,这表明 D. iridis nad3 基因曾经可能经过编辑。我们提出,D. iridis nad3 基因可能通过编辑转录本的逆转录而失去了对 RNA 编辑的需求,随后该转录本被插入基因组。