Sardu Alessandro, Treu Laura, Campanaro Stefano
Department of Biology, University of Padova, Via Ugo Bassi 58/b, 35131 Padova, Italy.
BMC Genomics. 2014 Dec 1;15(1):1045. doi: 10.1186/1471-2164-15-1045.
RNA-seq studies have an important role for both large-scale analysis of gene expression and for transcriptome reconstruction. However, the lack of software specifically developed for the analysis of the transcriptome structure in lower eukaryotes, has so far limited the comparative studies among different species and strains.
In order to fill this gap, an innovative software called ORA (Overlapped Reads Assembler) was developed. This software allows a simple and reliable analysis of the transcriptome structure in organisms with a low number of introns. It can also determine the size and the position of the untranslated regions (UTR) and of polycistronic transcripts. As a case study, we analyzed the transcriptional landscape of six S. cerevisiae strains in two different key steps of the fermentation process. This comparative analysis revealed differences in the UTR regions of transcripts. By extending the transcriptome analysis to yeast species belonging to the Saccharomyces genus, it was possible to examine the conservation level of unknown non-coding RNAs and their putative functional role.
By comparing the results obtained using ORA with previous studies and with the transcriptome structure determined with other software, it was proven that ORA has a remarkable reliability. The results obtained from the training set made it possible to detect the presence of transcripts with variable UTRs between S. cerevisiae strains. Finally, we propose a regulatory role for some non-coding transcripts conserved within the Saccharomyces genus and localized in the antisense strand to genes involved in meiosis and cell wall biosynthesis.
RNA测序研究对于基因表达的大规模分析和转录组重建都具有重要作用。然而,目前缺乏专门为分析低等真核生物转录组结构而开发的软件,这限制了不同物种和菌株之间的比较研究。
为了填补这一空白,开发了一种名为ORA(重叠读段组装器)的创新软件。该软件能够对内含子数量较少的生物体的转录组结构进行简单可靠的分析。它还可以确定非翻译区(UTR)和多顺反子转录本的大小和位置。作为案例研究,我们分析了六个酿酒酵母菌株在发酵过程的两个不同关键步骤中的转录情况。这种比较分析揭示了转录本UTR区域的差异。通过将转录组分析扩展到酿酒酵母属的酵母物种,可以研究未知非编码RNA的保守水平及其假定的功能作用。
通过将使用ORA获得的结果与先前的研究以及用其他软件确定的转录组结构进行比较,证明ORA具有很高的可靠性。从训练集中获得的结果使得能够检测酿酒酵母菌株之间具有可变UTR的转录本的存在。最后,我们提出了一些在酿酒酵母属内保守且位于参与减数分裂和细胞壁生物合成的基因反义链上的非编码转录本的调控作用。