Polysome Profiling and Metabolic Labeling Methods to Measure Translation in Trypanosoma brucei

Bajak, Kathrin; Clayton, Christine

doi:10.1007/978-1-0716-0294-2_7

Cited by 3 publications

(2 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nevertheless, the role of translational regulation of drug resistance in Leishmania has not been investigated yet. The major advantage of translational regulation is that it can be easily reversed and readjusted under different environments 63 . This study describes a new perspective on drug resistance in Leishmania parasites, highlighting translational reprogramming as a crucial driver of antimony-resistant phenotypes.…”

Section: Discussionmentioning

confidence: 99%

Translational reprogramming as a driver of antimony-drug resistance in Leishmania

et al. 2023

View full text Add to dashboard Cite

Leishmania is a unicellular protozoan that has a limited transcriptional control and mostly uses post-transcriptional regulation of gene expression, although the molecular mechanisms of the process are still poorly understood. Treatments of leishmaniasis, pathologies associated with Leishmania infections, are limited due to drug resistance. Here, we report dramatic differences in mRNA translation in antimony drug-resistant and sensitive strains at the full translatome level. The major differences (2431 differentially translated transcripts) were demonstrated in the absence of the drug pressure supporting that complex preemptive adaptations are needed to efficiently compensate for the loss of biological fitness once they are exposed to the antimony. In contrast, drug-resistant parasites exposed to antimony activated a highly selective translation of only 156 transcripts. This selective mRNA translation is associated with surface protein rearrangement, optimized energy metabolism, amastins upregulation, and improved antioxidant response. We propose a novel model that establishes translational control as a major driver of antimony-resistant phenotypes in Leishmania.

show abstract

Section: Discussionmentioning

confidence: 99%

Translational reprogramming as a driver of antimony-drug resistance in Leishmania

et al. 2023

View full text Add to dashboard Cite

show abstract

“…Polysome fractionation was performed according to Ref. 47. For each gradient, 5 3 10 8 cells were collected by centrifugation at 3000 rpm for 15 min.…”

Section: Polysome Fractionation and Analysismentioning

confidence: 99%

A potential role for a novel ZC3H5 complex in regulating mRNA translation in Trypanosoma brucei

Bajak

Leiss

Clayton

et al. 2020

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

In Trypanosoma brucei and related kinetoplastids, gene expression regulation occurs mostly post-transcriptionally. Consequently, RNA-binding proteins play a critical role in the regulation of mRNA and protein abundance. Yet, the roles of many RNA-binding proteins are not understood. Our previous research identified the RNA-binding protein ZC3H5 as possibly involved in gene repression, but its role in controlling gene expression was unknown. We here show that ZC3H5 is an essential cytoplasmic RNA-binding protein: RNAi targeting ZC3H5 causes accumulation of pre-cytokinetic cells followed by rapid cell death. Affinity purification and pair-wise yeast 2-hybrid analysis suggest that ZC3H5 forms a complex with three other proteins, encoded by genes Tb927.11.4900, Tb927.8.1500 and Tb927.7.3040. RNA immunoprecipitation revealed that ZC3H5 is preferentially associated with poorly translated, low-stability mRNAs, the 5'-untranslated regions and coding regions of which are enriched in the motif (U/A)UAG(U/A). As previously found in high-throughput analyses, artificial tethering of ZC3H5 to a reporter mRNA or other complex components repressed reporter expression. However, depletion of ZC3H5 in vivo caused only very minor decreases in a few targets, marked increases in the abundances of very stable mRNAs, an increase in monosomes at the expense of large polysomes, and appearance of "halfmer" disomes containing two 80S subunits and one 40S subunit. We speculate that the ZC3H5 complex might be implicated in quality control during the translation of sub-optimal open reading frames.

show abstract

A novel essential RNA-binding protein complex inTrypanosoma brucei

Bajak

Leiss

Clayton

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Polysome Profiling and Metabolic Labeling Methods to Measure Translation in Trypanosoma brucei

Cited by 3 publications

References 22 publications

Translational reprogramming as a driver of antimony-drug resistance in Leishmania

Translational reprogramming as a driver of antimony-drug resistance in Leishmania

A potential role for a novel ZC3H5 complex in regulating mRNA translation in Trypanosoma brucei

A novel essential RNA-binding protein complex inTrypanosoma brucei

Contact Info

Product

Resources

About