Pena Ana C, Pimentel Mafalda R, Manso Helena, Vaz-Drago Rita, Pinto-Neves Daniel, Aresta-Branco Francisco, Rijo-Ferreira Filipa, Guegan Fabien, Pedro Coelho Luis, Carmo-Fonseca Maria, Barbosa-Morais Nuno L, Figueiredo Luisa M
Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Av. Prof. Egas Moniz, Edifício Egas Moniz, 1649-028, Lisboa, Portugal.
Mol Microbiol. 2014 Aug;93(4):645-63. doi: 10.1111/mmi.12677. Epub 2014 Jul 14.
Trypanosoma brucei is a unicellular parasite that causes sleeping sickness in humans. Most of its transcription is constitutive and driven by RNA polymerase II. RNA polymerase I (Pol I) transcribes not only ribosomal RNA genes, but also protein-encoding genes, including variant surface glycoproteins (VSGs) and procyclins. In T. brucei, histone H1 (H1) is required for VSG silencing and chromatin condensation. However, whether H1 has a genome-wide role in transcription is unknown. Here, using RNA sequencing we show that H1 depletion changes the expression of a specific cohort of genes. Interestingly, the predominant effect is partial loss of silencing of Pol I loci, such as VSG and procyclin genes. Labelling of nascent transcripts with 4-thiouridine showed that H1 depletion does not alter the level of labelled Pol II transcripts. In contrast, the levels of 4sU-labelled Pol I transcripts were increased by two- to sixfold, suggesting that H1 preferentially blocks transcription at Pol I loci. Finally, we observed that parasites depleted of H1 grow almost normally in culture but they have a reduced fitness in mice, suggesting that H1 is important for host-pathogen interactions.
布氏锥虫是一种导致人类昏睡病的单细胞寄生虫。其大部分转录是组成型的,由RNA聚合酶II驱动。RNA聚合酶I(Pol I)不仅转录核糖体RNA基因,还转录蛋白质编码基因,包括可变表面糖蛋白(VSG)和前环素。在布氏锥虫中,组蛋白H1(H1)是VSG沉默和染色质浓缩所必需的。然而,H1在全基因组转录中是否发挥作用尚不清楚。在此,我们通过RNA测序表明,H1缺失会改变特定一组基因的表达。有趣的是,主要影响是Pol I基因座(如VSG和前环素基因)沉默的部分丧失。用4-硫尿苷标记新生转录本表明,H1缺失不会改变标记的Pol II转录本水平。相反,4sU标记的Pol I转录本水平增加了两到六倍,这表明H1优先阻断Pol I基因座的转录。最后,我们观察到缺失H1的寄生虫在培养中生长几乎正常,但它们在小鼠中的适应性降低,这表明H1对宿主-病原体相互作用很重要。