Transcriptomics

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Transcription profiling of T. brucei treated with dithiothreitol (DTT) or tunicamycin to invoke the unfolded protein response


ABSTRACT: Transcriptome responsiveness was further tested by attempts to invoke the unfolded protein repsonse (UPR), a classic ER-based pathway stimulated by the presence of increased levels of unfolded polypeptides. The UPR is mediated via transcriptional responses in both yeast and metazoan cells, and can be reliably activated by addition of dithiothreitol (DTT). Using DTT at concentrations that invoke a UPR in mammalian cells, Arabidopsis, yeast and other systems, we found that, in T. brucei, DTT exposure led to rapid cell death. We analysed the transcriptome at 1 mM DTT, under conditions where cells remained viable, as assessed by motility.

part 1: 3 biological replicates of SMB cells grown under normal conditions, and 3 replicates of SMB cells treated with 1mM DTT for 1hr, as well as dye swaps were used.

part 2: 3 biological replicates of SMB cells
grown under normal conditions, and 3 replicates of SMB cells treated
with 1mM DTT for 4hr, as well as dye swaps were used.

The UPR can also be activated by addition of tunicamycin. Using tunicamycin at concentrations that invoke a UPR in mammalian cells, Arabidopsis, yeast and other systems, we found that, in T. brucei, tunicamycin exposure efficiently inhibits trypanosome N-glycosylation and that it resulted in growth arrest over a period of up to 24 hours. We analysed the transcriptome at 5 ?g/ml tunicamycin under conditions where cells remained viable, as assessed by motility.

part 3: 3 biological replicates of SMB cells grown under normal conditions, and 3 replicates of SMB cells treated with 5 ?g/ml tunicamycin for 4hr, as well as dye swaps were used.

part 4: 3 biological replicates of SMB cells grown under normal conditions, and 3 replicates of SMB cells treated with 5 ?g/ml tunicamycin for 24hr, as well as dye swaps were used.

ORGANISM(S): Trypanosoma brucei  

SUBMITTER: Lila Koumandou  

PROVIDER: E-MEXP-1533 | ArrayExpress | 2008-06-26

REPOSITORIES: ArrayExpress

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Publications

The trypanosome transcriptome is remodelled during differentiation but displays limited responsiveness within life stages.

Koumandou V Lila VL   Natesan Senthil Kumar A SK   Sergeenko Tatiana T   Field Mark C MC  

BMC genomics 20080623


Trypanosomatids utilise polycistronic transcription for production of the vast majority of protein-coding mRNAs, which operates in the absence of gene-specific promoters. Resolution of nascent transcripts by polyadenylation and trans-splicing, together with specific rates of mRNA turnover, serve to generate steady state transcript levels that can differ in abundance across several orders of magnitude and can be developmentally regulated. We used a targeted oligonucleotide microarray, representin  ...[more]

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