Laboratorio de Parasitología Molecular, Departamento de Microbiología, Pontificia Universidad Javeriana, Bogotá, Colombia.
Parasitol Res. 2011 Mar;108(3):731-9. doi: 10.1007/s00436-010-2190-6. Epub 2010 Dec 4.
RNA editing in trypanosomatids is an elaborate form of post-transcriptional processing that inserts and deletes uridines in many mitochondrial pre-mRNAs, providing the genetic information needed to create functional transcripts. The process has been extensively analyzed in Trypanosoma brucei, Crithidia fasciculata, and Leishmania tarentolae. However, few data exist on this mechanism in pathogenic Leishmania species. Here, we show evidence that this process also operates in Leishmania braziliensis, being the first time that RNA editing has been described in a species of the Viannia subgenus. A partially edited transcript corresponding to the NADH dehydrogenase subunit 8 (ND8) gene was identified in L. braziliensis promastigotes. Sequence analysis allowed the identification of the maxicircle-encoded cryptogene, which shows a high degree of sequence conservation with the corresponding cryptogenes in other Leishmania species. Although an edition pattern could be postulated for the ND8 transcripts in L. braziliensis, attempts to isolate completely edited transcripts by RT-PCR were not fruitful; instead, many transcripts with partial and unexpected editing patterns were isolated. This data, together with our inability to detect full-size transcripts by Northern blotting in promastigotes of L. braziliensis, led us to the suggestion that the strain used in this study (M2904) lacks of critical RNA guides for a complete edition of ND8 transcripts.
RNA 编辑在动基体生物中是一种复杂的转录后加工形式,它在许多线粒体前体 mRNA 中插入和删除尿嘧啶,为创建功能性转录本提供了所需的遗传信息。该过程在布氏锥虫、福氏内格里虫和塔兰托利利什曼原虫中得到了广泛分析。然而,关于致病性利什曼原虫物种中的这种机制的数据很少。在这里,我们证明了这个过程也在巴西利什曼原虫中起作用,这是首次在 Viannia 亚属的一个物种中描述 RNA 编辑。在 L. braziliensis 前鞭毛体中鉴定到对应于 NADH 脱氢酶亚单位 8 (ND8) 基因的部分编辑转录本。序列分析允许鉴定出 maxicircle 编码的隐基因,该基因与其他利什曼原虫物种的相应隐基因具有高度的序列保守性。尽管可以推测 L. braziliensis 中的 ND8 转录本存在编辑模式,但通过 RT-PCR 分离完全编辑的转录本的尝试并不成功;相反,分离到许多具有部分和意外编辑模式的转录本。这些数据,以及我们在巴西利什曼原虫的前鞭毛体中通过 Northern blotting 无法检测到全长转录本,使我们推测在这项研究中使用的菌株 (M2904) 缺乏 ND8 转录本完全编辑所需的关键 RNA 指导。