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Nucleic Acids Res. 2014 Apr;42(6):3623-37. doi: 10.1093/nar/gkt1386. Epub 2014 Jan 17.

Comparative ribosome profiling reveals extensive translational complexity in different Trypanosoma brucei life cycle stages.

Author information

1
Research Center for Infectious Diseases, University of Wuerzburg, Wuerzburg 97080, Germany, Département Biologie cellulaire et infection, Institut Pasteur, Unité Biologie Cellulaire du Parasitisme, Paris 75015, France, INSERM U786, Paris 75015, France and Rudolf Virchow Center, University of Wuerzburg, Wuerzburg 97080, Germany.

Abstract

While gene expression is a fundamental and tightly controlled cellular process that is regulated at multiple steps, the exact contribution of each step remains unknown in any organism. The absence of transcription initiation regulation for RNA polymerase II in the protozoan parasite Trypanosoma brucei greatly simplifies the task of elucidating the contribution of translation to global gene expression. Therefore, we have sequenced ribosome-protected mRNA fragments in T. brucei, permitting the genome-wide analysis of RNA translation and translational efficiency. We find that the latter varies greatly between life cycle stages of the parasite and ∼100-fold between genes, thus contributing to gene expression to a similar extent as RNA stability. The ability to map ribosome positions at sub-codon resolution revealed extensive translation from upstream open reading frames located within 5' UTRs and enabled the identification of hundreds of previously un-annotated putative coding sequences (CDSs). Evaluation of existing proteomics and genome-wide RNAi data confirmed the translation of previously un-annotated CDSs and suggested an important role for >200 of those CDSs in parasite survival, especially in the form that is infective to mammals. Overall our data show that translational control plays a prevalent and important role in different parasite life cycle stages of T. brucei.

PMID:
24442674
PMCID:
PMC3973304
DOI:
10.1093/nar/gkt1386
[Indexed for MEDLINE]
Free PMC Article

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