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GNPS Proteomic Changes of Antimony Resistant Leishmania Growing Under Drug Challenge

ABSTRACT: Here, we used shotgun proteomics (LC-MS/MS) to identify changes in the proteome of antimony resistant parasites growing under drug challenge. We used a Leishmania tropica strain that was selected in vitro to resist high levels of trivalent antimony. Two experimental conditions were evaluated: a resistant strain grown without drug challenge (control), and a resistant strain grown under drug challenge (treatment). Three independent biological replicates were used per experimental condition.

INSTRUMENT(S): Q-Exactive Orbitrap

ORGANISM(S): Leishmania Tropica (ncbitaxon:5666)

SUBMITTER: Zemfira N. Karamysheva

PROVIDER: MSV000089617 | GNPS | Tue Jun 07 22:23:00 BST 2022

SECONDARY ACCESSION(S): PXD034401

REPOSITORIES: GNPS

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Project description:Purpose: In this study, we have used a translatomic approach by coupling polysome profiling and deep RNA-sequencing to estimate changes in the translatome of antimony-resistant Leishmania parasites Methods: Leishmania tropica promastigotes were stepwise selected for resistance to trivalent antimony. Two different strains were studied, the L. tropica SbIII-sensitive or wildtype strain (WT) and the derived highly resistant strain (HR). After polysome profiling four types of samples were evaluated by deep RNAseq: total mRNA used as input, monosomes (MS), light polysomes (LP), and heavy polysomes (HP). The DESeq2 algorithm was used for differential expression analysis to identify translational changes at the basal level (HR Vs. WT), translational changes to combat the drug (HR+SbIII Vs. HR), and to compare translatomic Vs. transcriptomic changes (HP Vs. Total input) [see overall design section below]. Results: Differential translational analysis (cutoff of fold change ≥ 1.5 and p-value corrected by Benjamini-Hochberg FDR ≤ 0.05) showed that transcripts composition per polysome fraction was different in the resistant strain. It included several upregulated (Up) and downregulated (Down) transcripts. At the basal level, 2431 different transcripts were differentially translated: monosome (Down: 4, Up: 0), light polysomes (Down: 906, Up: 951), and heavy polysomes (Down: 1096, Up: 1064). Under the antimony challenge, 189 different transcripts were differentially translated: monosome (Down: 0, Up: 2), light polysomes (Down: 9, Up: 57), and heavy polysomes (Down: 30, Up: 134). Overall, most of the changes were identified in polysome fraction when compared with monosomes or total transcriptome. Conclusions: Our study shows evidence that translational control has a main role in coordinating the resistance to antimony in Leishmania parasites. We propose a novel model that establishes translational control as a major driver of antimony-resistant phenotypes in Leishmania parasites.

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GNPS Proteomic Changes of Antimony Resistant Leishmania Growing Under Drug Challenge

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