Abstract
Messenger RNA is modified by the addition of a 5′ methylated cap structure, which protects the transcript and recruits protein complexes that mediate RNA processing and/or the initiation of translation. Two genes encoding mRNA cap methyltransferases have been identified in T. brucei: TbCMT1 and TbCGM1. Here we analysed the impact of TbCMT1 gene deletion on bloodstream form T. brucei cells. TbCMT1 was dispensable for parasite proliferation in in vitro culture. However, significantly decreased parasitemia was observed in mice inoculated with TbCMT1 null and conditional null cell lines. Using RNA-Seq, we observed that several cysteine peptidase mRNAs were downregulated in TbCMT1 null cells lines. The cysteine peptidase Cathepsin-L was also shown to be reduced at the protein level in TbCMT1 null cell lines. Our data suggest that TbCMT1 is not essential to bloodstream form T. brucei growth in vitro or in vivo but that it contributes significantly to parasite virulence in vivo.
Highlights
Trypanosoma brucei, a protozoan parasite transmitted by the tsetse fly, causes Human African Trypanosomiasis (HAT) and Nagana in cattle [1]
TbCMT1 is not required for Lister 427 bloodstream form (BSF) proliferation in vitro
We will refer to the single marker” (SM) cell line as wild type (WT) cells
Summary
Trypanosoma brucei, a protozoan parasite transmitted by the tsetse fly, causes Human African Trypanosomiasis (HAT) and Nagana in cattle [1]. When a tsetse fly feeds on infected human or animal blood, stumpy form T. brucei trypomastigotes enter the insect midgut and differentiate into the proliferating procyclic form (PCF) cells [2]. These migrate to the salivary glands and differentiate into proliferating epimastigote form cells and into non-dividing animal-infective metacyclic trypomastigotes [3]. During a subsequent blood meal, the tsetse fly transmits the metacyclic trypomastigotes into the hemolymphatic system of the host where they transform into the rapidly proliferating slender bloodstream form (BSF), and the cycle of infection begins again.
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