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Transcriptome analysis of Cymelarsan resistant and Cymelarsan sensitive strains of Trypanosoma brucei.

ABSTRACT: A drug sensitive stock of T.b.brucei (STIB247) was used to generate a drug resistant strain (247MelCyR), expressing 130-fold increase in resistance to the melaminophenyl arsenical Cymelarsan (MelCy) by subcurative treatment in mice. Expression of the Cymelarsan resistant phenotype was maintained in 247MelCyR following adaptation to procyclic form in vitro culture. Here we have compared the transcriptomes of the STIB247 and 247MelCyR strains from procyclic cell culture, using digital gene expression (DGE) analysis.

ORGANISM(S): Trypanosoma brucei brucei

PROVIDER: GSE23288 | GEO | 2011/07/28

SECONDARY ACCESSION(S): PRJNA131373

REPOSITORIES: GEO

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Transcriptome analysis of Cymelarsan resistant and Cymelarsan sensitive strains of Trypanosoma brucei.

Project description:A drug sensitive stock of T.b.brucei (STIB247) was used to generate a drug resistant strain (247MelCyR), expressing 130-fold increase in resistance to the melaminophenyl arsenical Cymelarsan (MelCy) by subcurative treatment in mice. Expression of the Cymelarsan resistant phenotype was maintained in 247MelCyR following adaptation to procyclic form in vitro culture. Here we have compared the transcriptomes of the STIB247 and 247MelCyR strains from procyclic cell culture, using digital gene expression (DGE) analysis. Digital gene expression analysis was performed on RNA derived from 2 biological replicates each of Cymelarsan resistant (247MelCyR) and Cymelarsan sensitive (STIB 247), T.b.brucei procyclic cell culture.

2011-07-28 | E-GEOD-23288 | biostudies-arrayexpress

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Project description:Trypanosome RNA polymerase II transcription is polycistronic, individual mRNAs being excised by trans splicing and polyadenylation. In this study, we refined the previously published mathematical model for bloodstream form parasites and extended it to the procyclic form. We used the model, together with known mRNA half-lives, to predict the abundances of individual mRNAs, assuming rapid, unregulated mRNA processing; then we compared the results with measured mRNA abundances. Remarkably, the abundances of most mRNAs in procyclic forms are predicted quite well by the model, being largely explained by variations in mRNA decay rates and length. In bloodstream forms substantially more mRNAs are less abundant than predicted. We list mRNAs that are likely to show particularly slow or inefficient processing, either in both forms or with developmental regulation. We also measured ribosome occupancies of all mRNAs in trypanosomes grown in the same conditions as were used to measure mRNA turnover. In procyclic forms there was a weak positive correlation between ribosome density and mRNA half-life, suggesting cross-talk between translation and mRNA decay; ribosome density was related to the proportion of the mRNA on polysomes, indicating control of translation initiation. Ribosomal protein mRNAs in procyclics appeared to be exceptionally rapidly processed but poorly translated. Through this study, we conclude that lLevels of mRNAs in procyclic form trypanosomes are determined mainly by length and mRNA decay, with some control of precursor processing. In bloodstream forms variations in nuclear events play a larger role in transcriptome regulation, suggesting acquisition of new control mechanisms during adaptation to mammalian parasitism.

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