in-vitro-differentiation-of-mature-myofibers-for-live-imaging

<em>In Vitro</em> Differentiation of Mature Myofibers for Live Imaging

Skeletal muscles are composed of myofibers, the biggest cells in the mammalian body and one of the few syncytia. How the complex and evolutionarily ...

Universidade de Lisboa

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.

Mafalda R. Pimentel

Instituto de Medicina Molecular,
Faculdade de Medicina,
Instituto de Medicina Molecular, Faculdade de Medicina

Universidade de Lisboa

In Vitro Differentiation of Mature Myofibers for Live Imaging

Mafalda R. Pimentel

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Universidade de Lisboa

In Vitro Differentiation of Mature Myofibers for Live Imaging

Instituto de Medicina Molecular,
Faculdade de Medicina,
Instituto de Medicina Molecular, Faculdade de Medicina