Biological Chemistry Department, Silberman Institute of Life Sciences, The Hebrew University of Jerusalem Jerusalem, Israel.
Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School Worcester, MA, USA.
Front Mol Neurosci. 2013 Dec 9;6:49. doi: 10.3389/fnmol.2013.00049. eCollection 2013.
Post-transcriptional control of gene expression has central importance during development and adulthood and in physiology in general. However, little is known about the extent of post-transcriptional control of gene expression in the brain. Most post-transcriptional regulatory effectors (e.g., miRNAs) destabilize target mRNAs by shortening their polyA tails. Hence, the fraction of a given mRNA that it is fully polyadenylated should correlate with its stability and serves as a good measure of post-transcriptional control. Here, we compared RNA-seq datasets from fly brains that were generated either from total (rRNA-depleted) or polyA-selected RNA. By doing this comparison we were able to compute a coefficient that measures the extent of post-transcriptional control for each brain-expressed mRNA. In agreement with current knowledge, we found that mRNAs encoding ribosomal proteins, metabolic enzymes, and housekeeping genes are among the transcripts with least post-transcriptional control, whereas mRNAs that are known to be highly unstable, like circadian mRNAs and mRNAs expressing synaptic proteins and proteins with neuronal functions, are under strong post-transcriptional control. Surprisingly, the latter group included many specific groups of genes relevant to brain function and behavior. In order to determine the importance of miRNAs in this regulation, we profiled miRNAs from fly brains using oligonucleotide microarrays. Surprisingly, we did not find a strong correlation between the expression levels of miRNAs in the brain and the stability of their target mRNAs; however, genes identified as highly regulated post-transcriptionally were strongly enriched for miRNA targets. This demonstrates a central role of miRNAs for modulating the levels and turnover of brain-specific mRNAs in the fly.
基因表达的转录后调控在发育和成年期以及一般生理学中具有核心重要性。然而,人们对大脑中基因表达的转录后调控程度知之甚少。大多数转录后调控效应因子(例如 miRNA)通过缩短其 polyA 尾巴使靶 mRNA 不稳定。因此,给定 mRNA 完全 polyA 化的部分应与其稳定性相关,并且是转录后控制的良好衡量标准。在这里,我们比较了来自果蝇大脑的 RNA-seq 数据集,这些数据集是从总 RNA(rRNA 耗尽)或 polyA 选择的 RNA 中生成的。通过进行这种比较,我们能够计算出一个衡量每个脑表达 mRNA 转录后控制程度的系数。与当前知识一致,我们发现编码核糖体蛋白、代谢酶和管家基因的 mRNAs 是转录后控制最少的 mRNA 之一,而那些已知高度不稳定的 mRNAs,如昼夜节律 mRNAs 和表达突触蛋白和具有神经元功能的蛋白质的 mRNAs,受到强烈的转录后控制。令人惊讶的是,后一组包括许多与大脑功能和行为相关的特定基因群。为了确定 miRNA 在这种调控中的重要性,我们使用寡核苷酸微阵列对果蝇大脑中的 miRNA 进行了分析。令人惊讶的是,我们没有发现 miRNA 在大脑中的表达水平与它们的靶 mRNA 稳定性之间存在很强的相关性;然而,被确定为高度转录后调控的基因强烈富集了 miRNA 靶标。这表明 miRNA 在调节果蝇大脑特异性 mRNAs 的水平和周转率方面发挥着核心作用。