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Transcriptomes of bloodstream-form trypanosomes from culture or buffy coat

ABSTRACT: Transcriptomes of bloodstream-form trypanosomes from culture or buffy coat

PROVIDER: PRJEB10042 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR973590.fastq.gz	Fastqsanger.gz
	ERR973591.fastq.gz	Fastqsanger.gz
	ERR973592.fastq.gz	Fastqsanger.gz
	ERR973593.fastq.gz	Fastqsanger.gz
	ERR973594.fastq.gz	Fastqsanger.gz

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PUF3 RNAi in bloodstream-form trypanosomes

Project description:PUF3 was depleted by RNAi for 24h in bloodstream-form trypanosomes (Trypanosoma brucei). No effect was seen on the transcriptome.

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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.

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Transcriptomes of bloodstream-form trypanosomes from culture or buffy coat

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Human buffy coat gene expression, custom 250-plex Nanostring panel

Project description:Buffy coat PBMC RNA expression was tested using a custom Nanostring probe panel, to identify general immune and glucocorticoid receptor-associated immune genes. ClinicalTrials.gov Identifier: NCT00824941

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DNA methylation profiles of human buffy coat samples from Huntington's disease

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

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Effect of ZC3H28 depletion (RNAi) in Trypanosoma brucei bloodstream forms

Project description:Bloodstream-form trypanosomes with inducible RNAi targeting ZC3H28 were incubated with the tetracycline inducer for 10h, cells without tetracycline were used as controls. This experiment was done in triplicates. In another similar experiment, bloodstream-form trypanosomes with inducible RNAi targeting ZC3H28 were incubated with the tetracycline inducer for 14h, 16h, and 24h. Cells without tetracycline (named ''no tet'' or ''0h'') were used as controls. This experiment was done in duplicates.

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Project description:Bloodstream form African trypanosomes are thought to rely exclusively upon glycolysis, using glucose as a substrate, for ATP production. Indeed, the pathway has long been considered a potential therapeutic target to tackle the devastating and neglected tropical diseases caused by these parasites. However, plasma membrane glucose and glycerol transporters are both expressed by trypanosomes and these parasites can infiltrate tissues that contain glycerol. Here, we show that bloodstream form trypanosomes can use glycerol for gluconeogenesis and for ATP production, particularly when deprived of glucose following hexose transporter depletion. We demonstrate that Trypanosoma brucei hexose transporters 1 and 2 (THT1 and THT2) are localized to the plasma membrane and that knockdown of THT1 expression leads to a growth defect that is more severe when THT2 is also knocked down. These data are consistent with THT1 and THT2 being the primary routes of glucose supply for the production of ATP by glycolysis. However, supplementation of the growth medium with glycerol substantially rescued the growth defect caused by THT1 and THT2 knockdown. Metabolomic analyses with heavy-isotope labelled glycerol demonstrated that trypanosomes take up glycerol and use it to synthesize intermediates of gluconeogenesis, including fructose 1,6-bisphosphate and hexose 6-phosphates, which feed the pentose phosphate pathway and variant surface glycoprotein biosynthesis. We used Cas9-mediated gene knockout to demonstrate a gluconeogenesis-specific, but fructose-1,6-bisphosphatase (Tb927.9.8720)-independent activity, converting fructose 1,6-bisphosphate into fructose 6-phosphate. In addition, we observed increased flux through the tricarboxylic acid cycle and the succinate shunt. Thus, contrary to prior thinking, gluconeogenesis can operate in bloodstream form T. brucei. This pathway, using glycerol as a physiological substrate, may be required in mammalian host tissues.

2018-12-10 | MTBLS706 | MetaboLights

Human buffy coat gene expression, custom 250-plex Nanostring panel

Project description:Human buffy coat gene expression, custom 250-plex Nanostring panel

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