German Primate Center, Gottingen, Germany.
J Toxicol Environ Health A. 2011;74(2-4):88-95. doi: 10.1080/15287394.2011.529057.
Editing of RNA molecules gained major interest when coding mRNA was analyzed. A small, noncoding, Alu DNA element transcript that may act as regulatory RNA in cells was examined in this study. Alu DNA element transcription was determined in buffy coat from healthy humans and human sporadic Creutzfeldt-Jakob disease (sCJD) cases. In addition, non-sCJD controls, mostly dementia cases and Alzheimer's disease (AD) cases, were included. The Alu cDNA sequences were aligned to genomic Alu DNA elements by database search. A comparison of best aligned Alu DNA sequences with our RNA/cDNA clones revealed editing by deamination by ADAR (adenosine deaminase acting on RNA) and APOBEC (apolipoprotein B editing complex). Nucleotide exchanges like a G instead of an A or a T instead of a C in our cDNA sequences versus genomic Alu DNA pointed to recent mutations. To confirm this, our Alu cDNA sequences were aligned not only to genomic human Alu DNA but also to the respective genomic DNA of the chimpanzee and rhesus. Enhanced ADAR correlated with A-G exchanges in dementia, AD, and sCJD was noted when compared to healthy controls as well as APOBEC-related C-T exchanges. The APOBEC-related mutations were higher in healthy controls than in cases suffering from neurodegeneration, with the exception of the dementia group with the prion protein gene (PRNP) MV genotype. Hence, this study may be considered the first real-time analysis of Alu DNA element transcripts with regard to editing of the respective Alu transcripts in human blood cells.
当对编码 mRNA 进行分析时,RNA 分子的编辑引起了广泛关注。本研究中检查了一种可能在细胞中作为调节 RNA 发挥作用的小的非编码 Alu DNA 元件转录本。在来自健康人类和人类散发性克雅氏病(sCJD)病例的白细胞层中测定 Alu DNA 元件转录。此外,还包括非 sCJD 对照物,主要是痴呆症病例和阿尔茨海默病(AD)病例。通过数据库搜索将 Alu cDNA 序列与基因组 Alu DNA 元件进行比对。将最佳比对的 Alu DNA 序列与我们的 RNA/cDNA 克隆进行比较,结果表明通过 ADAR(作用于 RNA 的腺苷脱氨酶)和 APOBEC(载脂蛋白 B 编辑复合物)脱氨酶进行编辑。与基因组 Alu DNA 相比,我们 cDNA 序列中的核苷酸交换,如 G 代替 A 或 T 代替 C,表明最近发生了突变。为了证实这一点,我们不仅将 Alu cDNA 序列与人类基因组 Alu DNA 进行比对,还与黑猩猩和恒河猴的相应基因组 DNA 进行比对。与健康对照组相比,在痴呆症、AD 和 sCJD 中增强的 ADAR 与 A-G 交换相关,与 APOBEC 相关的 C-T 交换相关。与患有神经退行性疾病的病例相比,健康对照组中的 APOBEC 相关突变更高,除了具有朊病毒蛋白基因(PRNP)MV 基因型的痴呆症组。因此,这项研究可以被认为是首次对人类血细胞中相应的 Alu 转录本进行实时分析,以研究 Alu DNA 元件转录本的编辑。