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Aggresomes protect mRNA under stress in Escherichia coli

ABSTRACT: To investigate the mechanism of mRNA protection in aggresomes, we used affinity purification mass spectrometry (AP-MS) to explore proteomes enriched or depleted in aggresomes.AP-MS analysis output in the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database revealed distinct protein-protein interaction (PPI) network topologies for aggresomes compared to the cytosol.

ORGANISM(S): Escherichia Coli

SUBMITTER: Yingying Pu

PROVIDER: PXD065481 | iProX | Wed Jun 25 00:00:00 GMT+01:00 2025

REPOSITORIES: iProX

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Aggresomes protect mRNA under stress in Escherichia coli

Project description:To investigate the mechanism of mRNA protection in aggresomes, we used affinity purification mass spectrometry (AP-MS) to explore proteomes enriched or depleted in aggresomes.

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Aggresomes protect mRNA under stress in Escherichia coli

Project description:Membraneless droplets formed through liquid-liquid phase separation of ribonucleoprotein particles contribute to mRNA storage in eukaryotic cells. How such aggresomes contribute to mRNA dynamics under stress, and their functional role, is less understood in bacteria. Here we used multiple approaches including imaging, modelling and transcriptomics to show that prolonged stress leading to ATP depletion in Escherichia coli results in increased aggresome formation, their compaction, and enrichment of mRNA within aggresomes compared to the cytosol. Transcript length was longer in aggresomes compared to the cytosol. Mass spectrometry showed exclusion of mRNA ribonucleases from aggresomes, which was due to negative charge repulsion. Experiments with fluorescent reporters and disruption of aggresome formation showed that mRNA storage within aggresomes promoted translation and associated with reduced lag phases during growth after stress removal. Our findings suggest that mRNA storage within aggresomes confers an advantage for bacterial survival and recovery from stress.

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Aggresomes protect mRNA under stress in Escherichia coli

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MULTI-TARGETED FLAVONOID INHIBITION OF THE PATHOGENIC BACTERIUM STAPHYLOCOCCUS AUREUS: A Proteomic Characterization

Project description:Growth inhibition of the pathogen Staphylococcus aureus with currently available antibiotics is problematic in part due to bacterial biofilm protection. While Although the recently characterized natural products, including 3´,4´,5-trihydroxy-6,7-dimethoxy-flavone [1], 3´,4´,5,6,7-pentahydroxy-flavone [2] and 5-hydroxy-4´,7-dimethoxy-flavone [3] exhibit both antibiotic and biofilm inhibitory activities, the mode of action of such hydroxylated flavonoids with respect to S. aureus inhibition has is yet to be characterized. Enzymatic digestion and high- resolution MS mass spectral analysis of differentially expressed proteins from S. aureus, with and without exposure to antibiotic flavonoids (1-3) has allowed for the characterization of global -protein alterations induced by metabolite treatment. A total of 56, 92, and 110 proteins are were differentially expressed with bacterial exposure to 1, 2 or 3, respectively. Connectivity of identified proteins is characterized using a search tool for the retrieval of interacting genes/proteins (STRING) with multi-targeted S. aureus inhibition of energy metabolism and biosynthesis by the assayed flavonoids observed. Identifying the mode of action of natural products as antibacterial agents is expected to provide insight into the potential use of flavonoids alone or in combination with known therapeutic agents to effectively control S. aureus infection.

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