Abstract
Arginine-Serine (RS) domain-containing proteins are RNA binding proteins with multiple functions in RNA metabolism. In mammalian cells this group of proteins is also implicated in regulation and coordination of cell cycle and apoptosis. In trypanosomes, an early branching group within the eukaryotic lineage, this group of proteins is represented by 3 members, two of them are SR proteins and have been recently shown to be involved in rRNA processing as well as in pre-mRNA splicing and stability. Here we report our findings on the 3rd member, the SR-related protein TbRRM1. In the present study, we showed that TbRRM1 ablation by RNA-interference in T. brucei procyclic cells leads to cell-cycle block, abnormal cell elongation compatible with the nozzle phenotype and cell death by an apoptosis-like mechanism. Our results expand the role of the trypanosomal RS-domain containing proteins in key cellular processes such as cell cycle and apoptosis-like death, roles also carried out by the mammalian SR proteins, and thus suggesting a conserved function in this phylogenetically conserved protein family.
Highlights
The protozoan parasite Trypanosoma brucei is the causative agent of sleeping sickness in humans, which remains a public health problem in sub-Saharan Africa, and the related disease “Nagana” in cattle
After 72 h post induction, cell viability was strongly compromised as shown by viability assays (S1 Fig). To demonstrate that this outcome was associated with a specific down regulation effect, TbRRM1 mRNA levels were determined by northern blot analysis
We show that TbRRM1 depletion induces growth arrest, morphological changes, aberrant N-K configurations, cell cycle block and cellular death by an apoptotic-like process in T. brucei procyclic cells (PC)
Summary
The protozoan parasite Trypanosoma brucei is the causative agent of sleeping sickness in humans, which remains a public health problem in sub-Saharan Africa, and the related disease “Nagana” in cattle. Sleeping sickness threatens millions of people in 36 countries, who live in remote areas with limited access to adequate health services, hampering the surveillance and the diagnosis and treatment of cases [1]. T. brucei belongs to the order Kinetoplastida and has a complex life cycle alternating between the tsetse fly and a mammalian host [2]. Its life cycle is characterized by a series of differentiation steps resulting in stages that differ morphologically, structurally and biochemically.
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