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Duration of the first steps of the human rRNA processing

机译:人类rRNA加工第一步的持续时间

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摘要

Processing of rRNA in mammalian cells includes a series of cleavages of the primary 47S transcript and results in producing three rRNAs: 18S, 28S and 5.8S. The sequence of the main processing events in human cells has been established, but little is yet known about the dynamics of this process, especially the dynamics of its early stages. In the present study, we used real-time PCR to measure levels of pre-rRNA after inhibition of transcription with actinomycin D. Thus we could estimate the half-life time of rRNA transcripts in two human-derived cell lines, HeLa and LEP (human embryonic fibroblasts), as well as in mouse NIH 3T3 cells. The primary transcripts seemed to be more stable in the human than in the murine cells. Remarkably, the graphs in all cases showed more or less pronounced lag phase, which may reflect preparatory events preceding the first cleavage of the pre-rRNA. Additionally, we followed the dynamics of the decay of the 5′ETS fragment which is degraded only after the formation of 41S rRNA. According to our estimates, the corresponding three (or four) steps of the processing in human cells take five to eight minutes.
机译:哺乳动物细胞中rRNA的加工包括一系列47S原始转录物的裂解,并导致产生三种rRNA:18S,28S和5.8S。已经确定了人类细胞中主要加工事件的顺序,但是对该过程的动力学,尤其是其早期阶段的动力学知之甚少。在本研究中,我们使用实时荧光定量PCR来检测放线菌素D抑制转录后的rRNA前水平。因此,我们可以估算出两种人类来源的细胞系HeLa和LEP中rRNA转录本的半衰期(人胚胎成纤维细胞)以及小鼠NIH 3T3细胞中。与人类细胞相比,人类的主要转录本似乎更稳定。值得注意的是,所有情况下的图均显示或多或少明显的滞后阶段,这可能反映了前rRNA第一次裂解之前的准备事件。此外,我们跟踪了仅在形成41S rRNA后才降解的5'ETS片段的衰减动力学。根据我们的估计,在人体细胞中进行相应的三个(或四个)处理过程需要五至八分钟。

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