Dataset Information

The copper-induced viable-but-non-culturable state in Cupriavidus metallidurans CH34 is characterized by distinct proteomic response patterns- Part III

ABSTRACT: Copper-based materials are actively explored for their potential as antimicrobial agents. However, recent studies show that sublethal concentrations of Cu ions can induce the viable-but-non-culturable (VBNC) cell state in certain bacteria, hampering with contamination control and monitoring. Despite the growing number of omics studies investigating the VBNC state, the underlying molecular mechanisms remain largely enigmatic. Here, we determine variations in protein expression of Cupriavidus metallidurans CH34 in different phases of the Cu-induced VBNC state via quantitative LC-MS/MS analysis at multiple sample time points. With this approach, we aimed to reveal cellular adaptations triggering VBNC formation and the characteristic spontaneous recovery of culturability (resuscitation). This submission contains controls and is part of the other pride submission PXD056297.

INSTRUMENT(S):

ORGANISM(S): Cupriavidus Metallidurans Ch34

SUBMITTER: Surya Gupta

LAB HEAD: Natalie Leys

PROVIDER: PXD056326 | Pride | 2025-05-07

REPOSITORIES: Pride

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Time-resolved proteomic profiling of Cupriavidus metallidurans CH34 in the copper-induced viable-but-nonculturable state.

Patocka Timothej T Gupta Surya S Mastroleo Felice F Leys Natalie N Matroule Jean-Yves JY Van Houdt Rob R

Metallomics : integrated biometal science 20250301 3

Copper-based materials are actively explored for their potential as antimicrobial agents. However, recent studies show that sublethal concentrations of Cu ions can induce the viable-but-nonculturable (VBNC) cell state in certain bacteria, hampering contamination control, and monitoring. In this study we contribute to the unravelling of this largely enigmatic phenomenon by determining the time-resolved proteome of Cu-treated Cupriavidus metallidurans CH34 during VBNC induction and resuscitation. ...[more]

PMID: 39963715

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Project description:Metabolically quiescent pathogens can persist in a viable non-replicating state for months or even years. In specific infectious diseases, such as tuberculosis, cryptococcosis, histoplasmosis, latent infection is a corollary of this dormant state, with at some point a phase of reactivation, leading to the symptoms of the infection. During murine cryptococcosis and macrophage uptake, stress and host immunity induce C. neoformans heterogeneity with generation of a sub-population of yeasts that has been shown to harbor a phenotype compatible with dormancy (low stress response, latency of growth). In this subpopulation, mitochondrial transcriptional activity has been shown to be up regulated and this phenotype has been considered as a hallmark of quiescence in stem cells. Based on these findings, we worked to reproduce this phenotype in vitro model and standardized the experimental conditions to generate dormancy in Cryptococcus neoformans. We found that incubation of stationary phase yeasts (STAT) in nutriment limited conditions and hypoxia during 8 days (8D-HYPOx) were able to mimic the phenotype obtained in vivo. In these conditions, mortality and/or apoptosis occurred in less than 5% of the yeasts compared to 30-40% of apoptotic or dead yeasts upon incubation in normoxia (8D-NORMOx). Yeasts in 8D-HYPOx harbored a lower stress response, delayed growth and less that 1% of culturability on agar plates, suggesting that these yeasts are viable but non culturable cells (VBNC). These VBNC were able to reactivate in the presence of pantothenic acid, a vitamin that is known to be involved in quorum sensing and precursor of acetyl-CoA. Global metabolism of 8D-HYPOx cells showed some specific requirements and was globally shut down compared to 8D-NORMOx and STAT conditions. Mitochondrial analyses showed that the mitochondrial masse increased with mitochondria mostly depolarized in 8D-HYPOx compared to 8D-NORMox with an increased expression of mitochondrial genes. Proteomic and transcriptomic analyses of 8D-HYPOx revealed that the number of secreted proteins and transcripts detected also decreased compared to 8D-NORMOx and STAT, but proteome, secretome and transcriptome harbored specific profiles that are engaged as soon as four days of incubation. Importantly, acetyl-CoA and fatty acid pathway involving mitochondria is required for the generation and viability maintenance of VBNC. All together, these data show that we were able to generate for the first time VBNC phenotype in Cryptococcocus neoformans. This VBNC state is associated with a reduced and specific metabolism that should be further studied to understand dormancy/quiescence in this organism.

			Action	DRS
	Control-1h-1.d.zip	Other
	Control-1h-2.d.zip	Other
	Control-1h-3.d.zip	Other
	Control-2h-1.d.zip	Other
	Control-2h-2.d.zip	Other