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The Long Noncoding RNA Transcriptome of Dictyostelium discoideum Development

机译:盘基网柄菌发育的长非编码RNA转录组。

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摘要

Dictyostelium discoideum live in the soil as single cells, engulfing bacteria and growing vegetatively. Upon starvation, tens of thousands of amoebae enter a developmental program that includes aggregation, multicellular differentiation, and sporulation. Major shifts across the protein-coding transcriptome accompany these developmental changes. However, no study has presented a global survey of long noncoding RNAs (ncRNAs) in D. discoideum. To characterize the antisense and long intergenic noncoding RNA (lncRNA) transcriptome, we analyzed previously published developmental time course samples using an RNA-sequencing (RNA-seq) library preparation method that selectively depletes ribosomal RNAs (rRNAs). We detected the accumulation of transcripts for 9833 protein-coding messenger RNAs (mRNAs), 621 lncRNAs, and 162 putative antisense RNAs (asRNAs). The noncoding RNAs were interspersed throughout the genome, and were distinct in expression level, length, and nucleotide composition. The noncoding transcriptome displayed a temporal profile similar to the coding transcriptome, with stages of gradual change interspersed with larger leaps. The transcription profiles of some noncoding RNAs were strongly correlated with known differentially expressed coding RNAs, hinting at a functional role for these molecules during development. Examining the mitochondrial transcriptome, we modeled two novel antisense transcripts. We applied yet another ribosomal depletion method to a subset of the samples to better retain transfer RNA (tRNA) transcripts. We observed polymorphisms in tRNA anticodons that suggested a post-transcriptional means by which D. discoideum compensates for codons missing in the genomic complement of tRNAs. We concluded that the prevalence and characteristics of long ncRNAs indicate that these molecules are relevant to the progression of molecular and cellular phenotypes during development.
机译:盘基网柄菌以单细胞的形式生活在土壤中,吞噬细菌并营养生长。饥饿后,成千上万的变形虫进入发育程序,其中包括聚集,多细胞分化和孢子形成。这些发育变化伴随着蛋白质编码转录组的重大转变。但是,尚无研究对盘实线虫中的长非编码RNA(ncRNA)进行过全面调查。为了表征反义和较长的基因间非编码RNA(lncRNA)转录组,我们使用选择性消除核糖体RNA(rRNA)的RNA测序(RNA-seq)文库制备方法分析了先前发表的发育时程样品。我们检测到了9833个蛋白质编码信使RNA(mRNA),621个lncRNA和162个假定的反义RNA(asRNA)的转录产物的积累。非编码RNA散布在整个基因组中,并且在表达水平,长度和核苷酸组成上各不相同。非编码转录组显示出与编码转录组相似的时间特征,逐渐变化的阶段散布着较大的飞跃。一些非编码RNA的转录谱与已知的差异表达编码RNA密切相关,暗示了这些分子在发育过程中的功能作用。检查线粒体转录组,我们建模了两个新颖的反义转录本。我们将另一种核糖体耗竭方法应用于部分样本,以更好地保留转移RNA(tRNA)转录本。我们观察到tRNA反密码子中的多态性,这表明D. discoideum通过一种转录后手段来补偿tRNA基因组补体中缺失的密码子。我们得出结论,长ncRNA的流行和特征表明这些分子与发育过程中分子和细胞表型的进展有关。

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