Rubin Carol M, Kimura Richard H, Schmid Carl W
Section of Molecular and Cellular Biology, University of California-Davis, Davis, CA 95616, USA.
Nucleic Acids Res. 2002 Jul 15;30(14):3253-61. doi: 10.1093/nar/gkf419.
Human Alu and adenovirus VA1 RNAs each stimulate the translational expression of reporter genes in co-transient transfection assays without affecting either the rate of global protein synthesis or the abundance of the reporter mRNA. This selective, post-transcriptional stimulation of expression, which is observed in human and mouse cell lines and for three reporters, acts through a PKR- independent mechanism. The activity of Alu and VA1 RNAs in this assay is transient, causing a reduction in the lag time for the translational expression of the newly synthesized reporter mRNAs. The reduction in this lag time accounts for the relative selectivity of the effect upon the expression of the reporter and suggests novel roles for Alu and VA1 RNA in cell stress recovery and viral infection. Deletion analysis demonstrates that a specific region residing within the right monomer of the dimeric Alu consensus sequence is necessary for activity. Highly abundant left Alu monomer transcripts are inactive but the right Alu monomer is fully active, although its transcripts are scarce. Mouse B1 and B2 SINE RNAs stimulate reporter gene expression in mouse cells, suggesting that this activity is a general property of eucaryotic SINEs.
在共瞬时转染实验中,人类Alu RNA和腺病毒VA1 RNA均可刺激报告基因的翻译表达,且不影响整体蛋白质合成速率或报告基因mRNA的丰度。这种在人类和小鼠细胞系中以及针对三种报告基因所观察到的对表达的选择性转录后刺激,是通过一种不依赖PKR的机制发挥作用的。在此实验中,Alu RNA和VA1 RNA的活性是短暂的,导致新合成的报告基因mRNA翻译表达的延迟时间缩短。这种延迟时间的缩短解释了对报告基因表达影响的相对选择性,并提示Alu RNA和VA1 RNA在细胞应激恢复和病毒感染中具有新的作用。缺失分析表明,二聚体Alu共有序列右单体中的一个特定区域对于活性是必需的。高度丰富的左Alu单体转录本无活性,但右Alu单体具有完全活性,尽管其转录本稀少。小鼠B1和B2短散在核元件(SINE)RNA可刺激小鼠细胞中的报告基因表达,表明这种活性是真核生物SINE的普遍特性。