Dataset Information

LC–MS/MS analysis of proteins co-immunoprecipitated with tagged PNO1 protein in Trypanosoma brucei

ABSTRACT: Tagging of endogenous PNO1 in Trypanosoma brucei was used to enable immunoaffinity purification of associated protein complexes. Co-purified proteins were analyzed by LC–MS/MS to identify ribosome biogenesis factors associatedwith PNO1

INSTRUMENT(S):

ORGANISM(S): Trypanosoma Brucei

SUBMITTER: Beatriz Guimarães

LAB HEAD: Nilson Zanchin

PROVIDER: PXD074183 | Pride | 2026-06-15

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	GFP-1_1_control_replicate1-run1.raw	Raw
	GFP-1_2_control_replicate1-run2.raw	Raw
	GFP-2_1_control_replicate2_run1.raw	Raw
	GFP-2_2_control_replicate2_run2.raw	Raw
	GFP-3_1_control_replicate3_run1.raw	Raw

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Publications

Proteomic Identification of Small-Subunit Ribosome Assembly Factors in <i>Trypanosoma brucei</i>.

Perez Gustavo Guadagnini GG Hiraiwa Priscila Mazzocchi PM da Silva Verônica Santana VS Zanchin Nilson Ivo Tonin NIT Guimarães Beatriz Gomes BG

Journal of proteome research 20260518 6

Trypanosomatid ribosomes display distinctive features, including extensive ribosomal RNA (rRNA) expansions and additional insertions in ribosomal proteins. Moreover, the region corresponding to the human 28S rRNA is fragmented into six molecules in these organisms with a duplication of the 3' fragment (ε) in<i>Leishmania</i>. Although these differences suggest that ribosome biogenesis in trypanosomatids may involve unique processing events, the molecular mechanisms underlying this process are st ...[more]

PMID: 42150138

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Project description:Variant Surface Glycoproteins (VSG) coat parasitic African trypanosomes and underpin antigenic variation and immune evasion. This VSG is a super-abundant virulence factor that is subject to post-transcriptional gene expression controls mediated via the VSG 3’-untranslated region (3’-UTR). To identify positive VSG regulators in bloodstream form cells, we used prior genome-scale screening data to prioritise mRNA binding protein (mRBPs) knockdowns that phenocopy VSG mRNA knockdown, displaying both loss-of-fitness and pre-cytokinesis accumulation. The top three candidate VSG regulators were CFB2 (cyclin F-box protein 2, Tb927.1.4650), MKT1 (Tb927.6.4770) and PBP1 (polyadenylate binding-protein binding-protein, Tb927.8.4540). Notably, CFB2 was recently found to regulate VSG transcript stability, and all three proteins were found to associate with each other. We used data-independent acquisition for accurate label-free quantification and deep proteome coverage to quantify expression profiles following depletion of each mRBP. Only CFB2 knockdown significantly reduced VSG expression and the expression of a reporter under the control of a VSG 3’-untranslated region (3’-UTR). CFB2 knockdown also triggered depletion of cytoplasmic ribosomal proteins, consistent with translation arrest observed when VSG synthesis is blocked. In contrast, PBP1 knockdown triggered depletion of CFB2, MKT1, and other components of the PBP1-complex. Finally, all three knockdowns triggered depletion of cytokinesis initiation factors, consistent with the cytokinesis defect that was confirmed here for all three knockdowns. Thus, genome-scale knockdown datasets facilitate the triage and prioritisation of candidate regulators. Quantitative proteomic analysis confirms 3’-UTR dependent positive control of VSG expression by CFB2 and interactions with additional mRBPs. Our results also reveal connections between VSG expression control by CFB2, ribosomal protein expression, and cytokinesis.

Dataset Information

LC–MS/MS analysis of proteins co-immunoprecipitated with tagged PNO1 protein in Trypanosoma brucei

Dataset's files

Publications

Proteomic Identification of Small-Subunit Ribosome Assembly Factors in <i>Trypanosoma brucei</i>.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

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Dataset Information

LC–MS/MS analysis of proteins co-immunoprecipitated with tagged PNO1 protein in Trypanosoma brucei

Dataset's files

Publications

Proteomic Identification of Small-Subunit Ribosome Assembly Factors in &lt;i&gt;Trypanosoma brucei&lt;/i&gt;.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Proteomic Identification of Small-Subunit Ribosome Assembly Factors in <i>Trypanosoma brucei</i>.