The English High School, Boston Public Schools, Boston, Massachusetts 02130, USA.
RNA. 2010 Apr;16(4):828-38. doi: 10.1261/rna.1989310. Epub 2010 Feb 16.
Regions of the Didymium iridis mitochondrial genome were identified with similarity to typical mitochondrial genes; however, these regions contained numerous stop codons. We used RT-PCR and DNA sequencing to determine whether, through RNA editing, these regions were transcribed into mRNAs that could encode functional proteins. Ten putative gene regions were examined: atp1, atp6, atp8, atp9, cox1, cox2, cytb, nad4L, nad6, and nad7. The cDNA sequences of each gene could encode a functional mitochondrial protein that was highly conserved compared with homologous genes. The type of editing events and editing sequence features were very similar to those observed in the homologous genes of Physarum polycephalum, though the actual editing locations showed a variable degree of conservation. Edited sites were compared with encoded sites in D. iridis and P. polycephalum for all 10 genes. Edited sequence for a portion of the cox1 gene was available for six myxomycetes, which, when compared, showed a high degree of conservation at the protein level. Different types of editing events showed varying degrees of site conservation with C-to-U base changes being the least conserved. Several aspects of single C insertion editing events led to the preferential creation of hydrophobic amino acid codons that may help to minimize adverse effects on the resulting protein structure.
鉴定了双色蜡蘑线粒体基因组的部分区域,这些区域与典型的线粒体基因具有相似性;然而,这些区域包含许多终止密码子。我们使用 RT-PCR 和 DNA 测序来确定,这些区域是否通过 RNA 编辑,转录成能够编码功能性蛋白质的 mRNAs。我们检查了 10 个假定的基因区域:atp1、atp6、atp8、atp9、cox1、cox2、cytb、nad4L、nad6 和 nad7。每个基因的 cDNA 序列都可以编码一个功能性的线粒体蛋白,与同源基因相比高度保守。编辑事件的类型和编辑序列特征与多头绒泡菌的同源基因非常相似,尽管实际的编辑位置显示出不同程度的保守性。编辑位点与双色蜡蘑和多头绒泡菌的编码位点进行了比较,涵盖了所有 10 个基因。cox1 基因部分的编辑序列可用于 6 种黏菌,比较后发现,在蛋白质水平上具有高度的保守性。不同类型的编辑事件在保守性方面存在差异,C 到 U 的碱基变化保守性最低。一些单 C 插入编辑事件的特征导致了疏水性氨基酸密码子的优先产生,这可能有助于最小化对产生的蛋白质结构的不利影响。