Pastro Lucía, Smircich Pablo, Di Paolo Andrés, Becco Lorena, Duhagon María A, Sotelo-Silveira José, Garat Beatriz
Laboratorio de Interacciones Moleculares, Facultad de Ciencias, Universidad de la RepúblicaMontevideo, Uruguay; Departamento de Genética, Facultad de Medicina, Universidad de la RepúblicaMontevideo, Uruguay.
Departamento de Genómica, Instituto de Investigaciones Biológicas Clemente Estable Montevideo, Uruguay.
Front Cell Dev Biol. 2017 Feb 13;5:8. doi: 10.3389/fcell.2017.00008. eCollection 2017.
In the protozoan parasite , as in other trypanosomatids, transcription of protein coding genes occurs in a constitutive fashion, producing large polycistronic transcription units. These units are composed of non-functionally related genes which are pervasively processed to yield each mRNA. Therefore, post-transcriptional processes are crucial to regulate gene expression. Considering that nuclear compartmentalization could contribute to gene expression regulation, we comparatively studied the nuclear, cytoplasmic and whole cell transcriptomes of the non-infective epimastigote stage of , using RNA-Seq. We found that the cytoplasmic transcriptome tightly correlates with the whole cell transcriptome and both equally correlate with the proteome. Nonetheless, 1,200 transcripts showed differential abundance between the nuclear and cytoplasmic fractions. For the genes with transcript content augmented in the nucleus, significant structural and compositional differences were found. The analysis of the reported epimastigote translatome and proteome, revealed scarce ribosome footprints and encoded proteins for them. Ontology analyses unveiled that many of these genes are distinctive of other parasite life-cycle stages. Finally, the relocalization of transcript abundance in the metacyclic trypomastigote infective stage was confirmed for specific genes. While gene expression is strongly dependent on transcript steady-state level, we here highlight the importance of the distribution of transcripts abundance between compartments in . Particularly, we show that nuclear compartmentation is playing an active role in the developmental stage determination preventing off-stage expression.
在原生动物寄生虫中,与其他锥虫一样,蛋白质编码基因的转录以组成型方式进行,产生大型多顺反子转录单元。这些单元由功能上不相关的基因组成,这些基因被广泛加工以产生每个mRNA。因此,转录后过程对于调节基因表达至关重要。考虑到核区室化可能有助于基因表达调控,我们使用RNA测序对的非感染性上鞭毛体阶段的核、细胞质和全细胞转录组进行了比较研究。我们发现细胞质转录组与全细胞转录组紧密相关,并且两者与蛋白质组的相关性相同。尽管如此,1200个转录本在核和细胞质部分之间显示出差异丰度。对于在细胞核中转录本含量增加的基因,发现了显著的结构和组成差异。对已报道的上鞭毛体翻译组和蛋白质组的分析揭示了稀少的核糖体足迹及其编码的蛋白质。本体分析表明,这些基因中的许多是其他寄生虫生命周期阶段所特有的。最后,证实了特定基因在循环后期锥鞭毛体感染阶段转录本丰度的重新定位。虽然基因表达强烈依赖于转录本稳态水平,但我们在此强调了转录本丰度在各部分之间分布在中的重要性。特别是,我们表明核区室化在发育阶段的确定中发挥着积极作用,防止阶段外表达。