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首页> 外文期刊>Journal of toxicology and environmental health, Part A >A comparative analysis to study editing of small noncoding BC200-and alu transcripts in brain of prion-inoculated rhesus monkeys (M. Mulatta)
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A comparative analysis to study editing of small noncoding BC200-and alu transcripts in brain of prion-inoculated rhesus monkeys (M. Mulatta)

机译：比较分析研究study病毒接种的恒河猴（M. Mulatta）脑中的小非编码BC200和alu转录本编辑的研究

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摘要

Small retroelements (short interspersed elements, abbreviated SINEs) are abundant in vertebrate genomes. Using RNA isolated from rhesus monkey cerebellum and buffy coat, reverse-transcription polymerase chain reaction (RT PCR) was applied to clone cDNA of BC200 and Alu RNAs. Transcripts containing Alu-SINE sequences may be subjected to extensive RNA editing by ADAR (adenosine deaminases that act on RNA) deamination. Abundance of Alu transcripts was determined with real-time RT PCR and was significantly higher than BC200 (brain cytoplasmic) in cerebellum. BC200 transcripts were absent from buffy coat cells. Availability of the rhesus genome sequence allowed the BC200 transcripts to be mapped to the specific locus on chromosome 13. Both the qualitative and quantitative characteristics of BC 200 expression argue for the BC 200 transcripts being generated by RNA polymerase III. In cerebellum, Alu transcripts often possessed base exchanges (A to G) consistent with ADAR editing and, somewhat unexpectedly, C to T exchanges consistent with APOBEC (apolipoprotein B editing complex) editing. In contrast, the BC200 transcripts, which as RNA POLIII transcripts play a role in dendritic RNA translation, appeared not to be deaminated, despite the presence of editing of Alu in the same tissue. To assess whether neuronal disease might influence editing of BC200 and Alu-SINE transcripts in cerebellum, RNA was isolated from two rhesus monkeys that were inoculated with prions from human variant Creutzfeldt-Jakob disease (vCJD). Regardless of prion-induced neurodegeneration, no BC200 RNA editing was observed, while Alu RNA continued to show both ADAR and APOBEC editing. Thus, BC200 RNAs do not appear to become accessible to editing enzymes despite infected neurons being subjected to severe stress, damage, and eventually cell death.
机译：脊椎动物基因组中存在少量的逆转录元素（短散布的元素，缩写为SINE）。使用从恒河猴小脑和血沉棕黄层分离的RNA，应用逆转录聚合酶链反应（RT PCR）克隆BC200和Alu RNA的cDNA。含有Alu-SINE序列的转录本可以通过ADAR（作用于RNA的腺苷脱氨基酶）脱氨作用进行广泛的RNA编辑。通过实时RT PCR测定Alu转录物的丰度，并显着高于小脑中的BC200（脑细胞质）。血沉棕黄层外壳细胞中不存在BC200转录本。恒河猴基因组序列的可用性使得BC200转录本可以定位到13号染色体上的特定基因座。BC 200表达的定性和定量特征都证明了RNA聚合酶III产生的BC 200转录本。在小脑中，Alu转录本通常具有与ADAR编辑一致的碱基交换（A到G），而出乎意料的是，与APOBEC（载脂蛋白B编辑复合体）编辑一致的C到T交换。相反，尽管在同一组织中存在Alu的编辑，但作为RNA POLIII转录物在树突状RNA翻译中起作用的BC200转录物似乎并未被脱氨基。为了评估神经元疾病是否会影响小脑中BC200和Alu-SINE转录本的编辑，从两只恒河猴中分离了RNA，这些猴子接种了人类变种Creutzfeldt-Jakob病（vCJD）的病毒。不论病毒引起的神经变性如何，均未观察到BC200 RNA编辑，而Alu RNA继续显示ADAR和APOBEC编辑。因此，尽管受感染的神经元受到了严重的压力，破坏，甚至最终导致细胞死亡，但BC200 RNA似乎仍无法被编辑酶所利用。

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