Wei Min, Zhao Xia, Liu Mi, Niu Meijuan, Seif Elias, Kleiman Lawrence
School of Medicine, Nankai University, Tianjin, China.
Lady Davis Institute, Jewish General Hospital, McGill University, Canada.
PLoS One. 2016 Apr 21;11(4):e0154044. doi: 10.1371/journal.pone.0154044. eCollection 2016.
In the current concept, tRNA maturation in vertebrate cells, including splicing of introns, trimming of 5' leader and 3' trailer, and adding of CCA, is thought to occur exclusively in the nucleus. Here we provide evidence to challenge this concept. Unspliced intron-containing precursor tRNAIle was identified in Human Immunodeficiency Virus type 1 (HIV-1) virions, which are synthesized in the cytoplasm. Northern blot, confocal microscopy and quantitative RT-PCR further verified enrichment of this unspliced tRNAIle within the cytoplasm in human cells. In addition to containing an intron, the cytoplasmic precursor tRNAIle also contains a short incompletely processed 5´ leader and a 3´ trailer, which abundance is around 1000 fold higher than the nuclear precursor tRNAIle with long 5' leader and long 3' trailer. In vitro data also suggest that the cytoplasmic unspliced end-immature precursor tRNAIle could be processed by short isoform of RNase Z, but not long isoform of RNase Z. These data suggest that precursor tRNAs could export from the nucleus to the cytoplasm in human cells, instead of be processed only in the nucleus.
在当前的概念中,脊椎动物细胞中的tRNA成熟,包括内含子的剪接、5'前导序列和3'尾随序列的修剪以及CCA的添加,被认为仅在细胞核中发生。在此,我们提供证据挑战这一概念。在细胞质中合成的1型人类免疫缺陷病毒(HIV-1)病毒粒子中鉴定出了含有未剪接内含子的前体tRNAIle。Northern印迹、共聚焦显微镜和定量RT-PCR进一步证实了这种未剪接的tRNAIle在人类细胞细胞质中的富集。除了含有一个内含子外,细胞质前体tRNAIle还含有一个短的未完全加工的5´前导序列和一个3´尾随序列,其丰度比具有长5'前导序列和长3'尾随序列的核前体tRNAIle高约1000倍。体外数据还表明,细胞质中未剪接的末端未成熟前体tRNAIle可被RNase Z的短同工型加工,但不能被RNase Z的长同工型加工。这些数据表明,前体tRNAs可以从细胞核输出到人类细胞的细胞质中,而不是仅在细胞核中进行加工。