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Trypannosome brucei brucei 427 TbTPR86 pullout

ABSTRACT: TPR86 is a tetra tricopeptide repeat containing protein, which we detected by the pull out experiment of TbSkp-Z, which involves in the regulation of surface proteome changes and suramin seisitivity. In this previous experiments, a deubiquitylase, USP7 was detected as a complex member with strong affinity. This is reverse pullout to confirm the TUS (TPR86, USP7, SKP-Z) complex members.

INSTRUMENT(S):

ORGANISM(S): Trypanosoma Brucei

DISEASE(S): Trypanosomiasis

SUBMITTER: Kayo Ono

LAB HEAD: Mark C Field

PROVIDER: PXD021797 | Pride | 2025-05-06

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	KO_TP1.raw	Raw
	KO_TP2.raw	Raw
	KO_TP3.raw	Raw
	KO_WT1.raw	Raw
	KO_WT2.raw	Raw

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Publications

TUSK: a ubiquitin hydrolase complex modulating surface protein abundance in trypanosomes.

Yamada Kayo K Zhang Ning N Yaqub Farzana K FK Zoltner Martin M Field Mark C MC

Frontiers in parasitology 20230427

Control of protein levels is vital to cellular homeostasis, for maintaining a steady state, to coordinate changes during differentiation and other roles. In African trypanosomes surface proteins contribute to immune evasion, drug sensitivity and environmental sensing. The trypanosome surface is dominated by the GPI-anchored variant surface glycoprotein, but additional GPI-anchored and <i>trans</i>-membrane domain proteins are present with known roles as nutrient receptors and signal transducers. ...[more]

PMID: 39816833

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Trypanosoma brucei global proteome changes upon suramin exposure

Project description:To understand how suramin kills Trypanosoma brucei, we examined the impact on proteome at different timepoints

2020-04-30 | PXD014215 | Pride

The endosomal TbTpr86/TbUsp7/SkpZ (TUS) complex controls surface protein abundance in trypanosomes

Project description:In trypanosomes two deubiquitilating enzymes (DUBs), orthologous to human USP7 and VDU1, control abundance of a cohort of surface proteins, including invariant surface glycoproteins (ISGs). Silencing TbUsp7 partially inhibits endocytosis and invariant surface glycoprotein turnover. S-phase kinase-associated protein 1 (Skp1) has crucial roles in cell cycle progression, transcriptional regulation, signal transduction and other processes in animals and fungi by virtue of being a component of cullin E3 ubiquitin ligases. Unexpectedly, trypanosomes possess multiple Skp1 paralogs, including a divergent paralog designated SkpZ. SkpZ is implicated in suramin-sensitivity and endocytosis and decreases in abundance following TbUsp7 knockdown. SkpZ physically interacts with TbUsp7 and TbTpr86. The latter is a tetratricopeptide-repeat protein also implicated in suramin sensitivity and located close to the flagellar pocket/endosomes, consistent with a role in endocytosis. Further, silencing SkpZ reduced abundance of TbUsp7 and TbTpr86 and many trans-membrane domain surface proteins. Our data indicate that TbTpr, TbUsp7 and SkpZ form the ‘TUS’ complex that regulates abundance of a significant cohort of trypanosome surface proteins.

2025-01-08 | PXD021000 | Pride

Whole cell proteomics analysis of Trypanosoma brucei 75kDa invariant surface glycoprotein knockout cells

Project description:A superfamily of invariant surface glycoproteins (ISGs) populates the African trypanosome surface, one of which, ISG75, is implicated in uptake of the century-old drug suramin. We disrupted the ISG75 gene locus encoding an array of six ISG75 paralogs on chromosome V by CRISPR/Cas9 knockout. Here we characterise this knockout strain by quantitative whole cell proteomics. Additionally, we analyse the way ISG75 knockout cells respond to suramin on the proteome level.

2023-03-11 | PXD031887 | Pride

USP7 is a tumor-specific WNT activator for APC-mutated colorectal cancer by mediating B-catenin deu

Project description:The tumor suppressor gene adenomatous polyposis coli (APC) is mutated in most colorectal cancers (CRC) resulting in constitutive Wnt activation. To understand the Wnt-activating mechanism of APC mutation, we applied CRISPR/Cas9 technology to engineer various APC-truncated isogenic lines. We find that the β-catenin inhibitory domain (CID) in APC represents the threshold for pathological levels of Wnt activation and tumor transformation. Mechanistically, CID-deleted APC truncation promotes β-catenin deubiquitination through reverse binding of β-TrCP and USP7 to the destruction complex. USP7 depletion in APC-mutated CRC inhibits Wnt activation by restoring β-catenin ubiquitination, drives differentiation and suppresses xenograft tumor growth. Finally, the Wnt-activating role of USP7 is specific to APC mutations, thus can be used as tumor-specific therapeutic target for most CRCs.

2017-11-09 | PXD007947 | Pride

Trypanosoma brucei rhodensience suramin resistance shotgun proteomics

Project description:Human African Trypanosomiasis (HAT) is a disease of major economic importance in Sub-Saharan Africa. In eastern and southern Africa. Here we analysed clinical isolates of T brucei rhodensiense, resistant to suramin by shotgun proteomics . And identified parasite proteins whose expression is associated with resistance to suramin.

2021-09-09 | PXD021560 | Pride

Transcriptomics of suramin resistance in Trypanosoma brucei

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The nuclear proteome of Trypanosoma brucei

Project description:Trypanosoma brucei is a protozoan flagellate that is transmitted by tsetse flies into the mammalian bloodstream. The parasite has a huge impact on human health both directly by causing African sleeping sickness and indirectly, by infecting domestic cattle. The biology of trypanosomes involves some highly unusual, nuclear-localised processes. These include polycistronic transcription without classical promoters initiated from regions defined by histone variants, trans-splicing of all transcripts to the exon of a spliced leader RNA, transcription of some very abundant proteins by RNA polymerase I and antigenic variation, a switch in expression of the cell surface protein variants that allows the parasite to resist the immune system of its mammalian host. Here, we provide the nuclear proteome of procyclic Trypanosoma brucei, the stage that resides within the tsetse fly midgut. We have performed quantitative label-free mass spectrometry to score 764 significantly nuclear enriched proteins in comparison to whole cell lysates. A comparison with proteomes of several experimentally characterised nuclear and non-nuclear structures and pathways confirmed the high quality of the dataset: the proteome contains about 80% of all nuclear proteins and less than 2% false positives. Using motif enrichment, we found the amino acid sequence KRxR present in a large number of nuclear proteins. KRxR is a sub-motif of a classical eukaryotic monopartite nuclear localisation signal and could be responsible for nuclear localization of proteins in Kinetoplastida species. As a proof of principle, we have confirmed the nuclear localisation of six proteins with previously unknown localisation by expressing eYFP fusion proteins. While proteome data of several T. brucei organelles have been published, our nuclear proteome closes an important gap in knowledge to study trypanosome biology, in particular nuclear-related processes.

2017-07-31 | PXD006745 | Pride

USP7 cooperates with SCML2 to regulate the activity of PRC1

Project description:In this experiment, we sought to identify the distribution of USP7 on chromatin relative to other PRC1 components ChIP-seq for USP7 in 293T-REx cells

2015-06-10 | E-GEOD-61048 | biostudies-arrayexpress

Characterisation of cullin E3 ubiquitin ligases in trypanosomes

Project description:Trypanosomes have seven cullin paralogs, the majority lineage-specific. Six of those cullins (TbCul-A, TbCul-C, TbCul-D TbCul-E, TbCul-F and TbCul-G) were endogenously 3xHA-epitope tagged at the C-terminus. We used affinity capture/mass spectrometry to determine the composition of these cullin complexes. TbCul-A and TbCul-E function were further studied by analysis of proteome changes upon upon RNAi.

2024-03-25 | PXD042022 | Pride

Microarry from Treg with conditional knockout of Usp7

Project description:Microarry from Treg with conditional knockout of Usp7 RNA from three independent samples from FACS sorted CD4+YFP+ Treg of fl-Usp7/Foxp3cre mice, compared to Foxp3cre control (C57BL/6 background).

2015-09-15 | E-GEOD-72988 | biostudies-arrayexpress

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