Dataset Information

Translational control by Trypanosoma brucei DRBD18 contributes to the maintenance of the procyclic state

ABSTRACT: Trypanosoma brucei occupies distinct niches throughout its life cycle, both within the vertebrate host (fat tissue, bloodstream, central nervous system) and within the insect host, the tsetse fly (taken up with a bloodmeal and passing through the midgut, and migrating to the salivary glands). The immunological and biochemical complexity and variability of each of these environments require a reshaping of the protein landscape of the parasite both to evade surveillance and face changing metabolic demands. Whereas most well-studied organisms rely on transcriptional control as the main regulator of gene expression, post-transcriptional control mechanisms are particularly important in T. brucei, and these are often mediated by RNA-binding proteins. DRBD18 is a T. brucei RNA binding protein that interacts with ribosomal proteins and translation factors. Here, we tested a role for DRBD18 in translational control. We show that DRBD18 depletion by RNA interference leads to altered polysomal profiles with a specific depletion of heavy polysomes. RiboSeq analysis reveals that 101 transcripts change in translational efficiency (TE) upon DRBD18 depletion: 41 of them exhibit decreased TE and 60 increased TE. A further 66 transcripts are buffered, i.e. changes in transcript abundance are compensated by changes in TE such that the total translational output is expected not to change. Proteomic analysis validates much of these data. In DRBD18 depleted cells, a cohort of transcripts that code for procyclic form specific proteins are translationally repressed while, conversely, transcripts that code for bloodstream form specific proteins are translationally enhanced. These data suggest that DRBD18 contributes to the maintenance of the procyclic state through both positive and negative translational control of specific mRNAs.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Trypanosoma Brucei

SUBMITTER: Shichen Shen

LAB HEAD: Jun Qu

PROVIDER: PXD039064 | Pride | 2024-01-26

REPOSITORIES: Pride

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			Action	DRS
	20220818_LRead_R1DOX_240K_clm203_WC18_63cm_HCD_IT_AGtrap_val50_9to30B_180m_4ul_01.msf	Msf
	20220818_LRead_R1DOX_240K_clm203_WC18_63cm_HCD_IT_AGtrap_val50_9to30B_180m_4ul_01.raw	Raw
	20220818_LRead_R1_240K_clm203_WC18_63cm_HCD_OT_AGtrap_val50_9to30B_180m_4ul_01.msf	Msf
	20220818_LRead_R1_240K_clm203_WC18_63cm_HCD_OT_AGtrap_val50_9to30B_180m_4ul_01.raw	Raw
	20220818_LRead_R1_240K_clm203_WC18_63cm_HCD_OT_AGtrap_val50_9to30B_180m_4ul_01_20220819143155.msf	Msf

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Translational control by Trypanosoma brucei DRBD18 contributes to the maintenance of the procyclic state.

Ciganda Martin M Sotelo-Silveira José J Dubey Ashutosh P AP Pandey Parul P Smith Joseph T JT Shen Shichen S Qu Jun J Smircich Pablo P Read Laurie K LK

RNA (New York, N.Y.) 20230920 12

Trypanosoma brucei occupies distinct niches throughout its life cycle, within both the mammalian and tsetse fly hosts. The immunological and biochemical complexity and variability of each of these environments require a reshaping of the protein landscape of the parasite both to evade surveillance and face changing metabolic demands. In kinetoplastid protozoa, including T. brucei, posttranscriptional control mechanisms are the primary means of gene regulation, and these are often me ...[more]

PMID: 37730435

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Project description:Trypanosoma brucei subspecies infect humans and animals in sub-Saharan Africa. This early diverging eukaryote shows many novel features in basic biological processes, including the use of polycistronic transcription to generate all protein-coding mRNAs. Therefore we hypothesized that translational control provides a means to tune gene expression during parasite development in mammalian and fly hosts. We used ribosome profiling to examine genome-wide protein production in animal-derived slender bloodstream forms and cultured procyclic (insect midgut) forms. About one-third of all CDSs showed statistically significant regulation of protein production between the two stages. Of these, more than two-thirds showed a change in translation efficiency, but few were controlled by this parameter alone. Ribosomal proteins were translated poorly, especially in animal-derived parasites. A disproportionate number of metabolic enzymes were up-regulated at the mRNA level in procyclic forms, as were variant surface glycoproteins in bloodstream forms. Comparison with cultured bloodstream forms from another strain identified stage-specific changes in protein production that transcend strain and growth conditions. Genes with upstream ORFs had a lower mean translation efficiency but no evidence was seen for involvement of these ORFs in stage-regulation. Ribosome profiling revealed that differences in the production of specific proteins in T. brucei slender bloodstream and procyclic forms are more common than anticipated from analysis of mRNA abundance. While in vivo and in vitro derived slender bloodstream forms of different strains are more similar to one another than to procyclic forms, they showed numerous differences at both the mRNA and protein production level. Ribosome profiling and mRNA libraries were constructed in triplicate from in vitro PCF and in vivo BF lifestages of theT. brucei Treu927 and in vitro T. brucei Lister427, to evaluate role of translational gene regulation

Dataset Information

Translational control by Trypanosoma brucei DRBD18 contributes to the maintenance of the procyclic state

Dataset's files

Publications

Translational control by <i>Trypanosoma brucei</i> DRBD18 contributes to the maintenance of the procyclic state.

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