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Trypanosoma brucei whole cell proteome changes upon exposure to the aminomethyl-benzoxaborole AN3057

ABSTRACT: Benzoxaboroles (BoBs) feature a boron-heterocyclic core and are an important recent innovation in the development of drugs against a range of pathogens and other pathologies. A broad spectrum of pharmacology is associated with chemically diverse BoB derivatives and includes multiple modes-of-action and targets. However, a consensus MoA for BoBs targeting evolutionarily diverse protozoan pathogens has emerged with the identification of CPSF3/CPSF73 in the CPSF complex in both apicomplexan and kinetoplastida parasites. We have detected a functional connection between protein sumoylation and the BoB boron-heterocyclic scaffold using comprehensive genetic screens in Trypanosoma brucei. Strikingly, as part of this sumoylation response, members of the CPSF complex are specifically and rapidly destabilised in a SUMO and proteosome-dependent manner. Here we deposit SILAC proteomics data quantifying the effects of BoB exposure on the global protein landscape in T. brucei.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Trypanosoma Brucei

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Trypanosomiasis

SUBMITTER: Martin Zoltner

LAB HEAD: Dr. Martin Zoltner

PROVIDER: PXD049328 | Pride | 2024-03-25

REPOSITORIES: Pride

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Project description:Endometrial cancer (EC) is the most common cancer of female reproductive organs. Because some low-grade ECs might also experience tumor recurrence after surgery and a worse prognosis, the study of alterations related to EC pathogenesis of the disease might help to get insights into underlying mechanisms involved in EC development and metastasis and identify novel markers associated to the disease. Here, low C1GALT1 protein expression levels were associated to a more aggressive phenotype of EC. Then, we aimed at investigating the role of C1GALT1 in EC progression by quantitative proteomics. ECC-1 cells were used as endometrioid EC model, and the effect of C1GALT1 depletion was analyzed using C1GALT1 specific short hairpin RNAs (shRNA) in comparison to SCRAMBLE shRNA. SILAC and mass spectrometry analysis were performed to identify and quantify dysregulated proteins associated with C1GALT1 depletion in the cell extract and secretome proteome of shC1GALT1 and SCRAMBLE ECC-1 cells. Out of 5208 proteins identified and quantified by LC-MS/MS, 76 and 143 proteins showed dysregulation (fold-change ≥1.5 or ≤0.67) in shC1GALT1 ECC-1 cells’ extracts and secretome, respectively. Nine dysregulated proteins were selected for validation by orthogonal techniques, confirming their dysregulation upon C1GALT1 depletion. Bioinformatics analyses pointed out to an increase in pathways and dysregulated proteins that associated with more aggressive phenotype. This finding was corroborated by loss-of-function cell-based assays. A higher proliferation, invasion, migration, colony formation and angiogenesis capacity of C1GALT1 depleted cells was observed. Finally, the negative protein expression correlation found by proteomics between C1GALT1 and LGALS3 was confirmed by IHC in actual EC samples, suggesting C1GALT1 stably depleted ECC-1 cells mimic the aggressive phenotype of EC cells and might be useful for the identification and validation of potential markers in aggressive ECs.

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Trypanosoma brucei whole cell proteome changes upon exposure to the aminomethyl-benzoxaborole AN3057

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