Project description:It has been shown that in mammalian cells alternative transcription initiation is extensively regulated during development and across cell-types, which confers dynamic transcript 5âUTR repertoire. However it is underexplored how the heterogeneity of 5âUTR isoforms would affect the downstream steps for protein expression, such as translation. To this end, we globally compared the translational profile of distinct mRNA TSS isoforms in mouse fibroblast cells, by combining deep-sequencing based mRNA 5âends profiling and polysome fractionation. We demonstrated the extensive translation regulation conferred by TSS heterogeneity. 5'end sequencing in seven polysome fractions, in two replicates, using Illumina Hiseq2000

2016-07-20 | E-GEOD-78241 | biostudies-arrayexpress

Quantitative proteomics upon CARP3 knock down in Trypanosoma brucei bloodstream forms

Project description:How flagellar signaling regulates the host interaction of parasites remains a challenge due to poor conservation of signaling systems with those in cilia of higher organisms. The trypanosome-specific cAMP response protein 3 (CARP3) shows developmentally regulated localization at the flagellar tip membrane, where it is essential for parasite swarming and colonization of the tsetse fly insect vector. This project describes a label-free, quantitative proteomics approach that identifies proteins changing in abundance upon inducible CARP3 knock down in bloodstream stage Trypanosoma brucei.

2022-08-31 | PXD025401 | Pride

Pull-down of PTP-tagged EIF4E2 from bloodstream-form Trypanosoma brucei

Project description:Pull-down of PTP-tagged EIF4E2 from bloodstream-form Trypanosoma brucei, sequence of bound and unbound fractions

2021-08-31 | E-MTAB-10670 | biostudies-arrayexpress

Heat shock of procyclic Trypanosoma brucei.

Project description:Polysome gradients on Trypanosoma brucei grown at 27C or after 1 h at 39C

2016-06-13 | E-MTAB-4555 | biostudies-arrayexpress

Translation profiling of Schizosaccharomyces pombe size separated polysome fractions provides data on average numbers of associated ribosomes for most transcripts

Project description:To obtain translational profiles for all mRNAs, polysome preparations are separated according to their size using a sucrose gradient and the mRNAs in each fraction are identified and quantified with DNA microarrays. Starting with exponentially growing cells, we analyzed 12 polysome fractions using DNA microarrays containing elements for all known and predicted genes of fission yeast. This approach provided data on average numbers of associated ribosomes for most transcripts.

2007-03-22 | E-TABM-222 | biostudies-arrayexpress

TbSAP is a novel chromatin protein repressing metacyclic Variant Surface Glycoprotein expression sites in bloodstream form Trypanosoma brucei

Project description:The African trypanosome Trypanosoma brucei is a unicellular eukaryote, which relies on a protective Variant Surface Glycoprotein (VSG) coat for survival in the mammalian host. A single trypanosome has >2000 VSG genes and pseudogenes of which only one is expressed from one of ~15 telomeric bloodstream form expression sites (BESs). Infectious metacyclic trypanosomes present within the tsetse fly vector also express VSG from a separate set of telomeric metacyclic ESs (MESs). All MESs are silenced in bloodstream form T. brucei. As very little is known about how this is mediated, we performed a whole genome RNAi library screen to identify MES repressors. This allowed us to identify a novel SAP domain containing DNA binding protein which we called TbSAP. TbSAP is enriched at the nuclear periphery and binds both MESs and BESs. Knockdown of TbSAP in bloodstream form trypanosomes did not result in cells becoming more ‘metacyclic’-like. Instead, there was extensive global upregulation of transcripts including MES VSGs, VSGs within the silent VSG arrays as well as genes immediately downstream of BES promoters. TbSAP therefore appears to be a novel architectural chromatin protein playing an important role in silencing the extensive VSG repertoire of bloodstream form T. brucei.

2020-11-04 | GSE160713 | GEO

Polysome-seq in zebrafish early embryonic development

Project description:We followed the polysomal association of maternal and early zygotic transcriptome over the first few hours of embryonic development, prior to and after MBT. We isolated polysome-associated (bound) and non-polysome-associated (unbound) mRNAs using sucrose gradient centrifugation followed by size fractionation. Using next generation sequencing (RNA-seq), we profiled the transcriptome in polysome-bound and unbound fractions. Our analysis revealed distinct dynamics of polysome association of cytoplasmically polyadenylated maternal mRNAs.

2017-10-14 | GSE88776 | GEO

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