Dataset Information

Sulfur Starvation Induces an Fe-Replete Response and Attenuates Virulence Pathways in Pseudomonas aeruginosa PAO1

ABSTRACT: Specific bacteria grown with alternative sulfur sources leads to metabolic reprogramming that includes the expression of sulfur-scavenging and antioxidant genes. To determine the detailed metabolic effect of sulfur starvation, P. aeruginosa PAO1 was grown in sulfur-free media alone and with the addition of sulfate and evaluated by label-free proteomics. Proteomics analyses identified 729 differentially expressed proteins from 2633 confidently identified proteins. Differentially expressed features were analyzed for pathway enrichment using STRING (proteomics),leveraging Gene Ontology and KEGG annotations. corresponding proteins involved in sulfur assimilation and metabolism, including those of the msu, ssu, and cys operons, showed an increased abundance during sulfur starvation. Sulfur starvation depressed Fe-uptake gene/pathway expression, including the pyoverdine (pvd), pyochelin (pch), and heme-uptake (phu and has) operons. Expression of genes from virulence pathways was also diminished, including quorum sensing, MDR efflux pumps, phenazine biosynthetic genes, and hydrolytic enzymes thought to be regulated by the ferric uptake regulator and PrrF sRNAs. In contrast, there was a significant increase in expression of proteins involved in Fe-storage, including bfrB, dps, and PA4880. Our study demonstrates that sulfur starvation leads to an Fe-replete response in P. aeruginosa PAO1 due to a disruption in Fe homeostasis and uncovered the suppression of critical virulence gene products during sulfur starvation.

INSTRUMENT(S):

ORGANISM(S): Pseudomonas Aeruginosa Atcc 700888 Bacteria

SUBMITTER: Tonya Zeczycki

LAB HEAD: Holly R. Ellis

PROVIDER: PXD062994 | Pride | 2025-11-10

REPOSITORIES: Pride

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Sulfur starvation induces an Fe-replete response and attenuates virulence pathways in Pseudomonas aeruginosa PAO1.

Ugochukwu Chidozie G CG Schwartz Tonia S TS Zeczycki Tonya N TN Goodwin Douglas C DC Ellis Holly R HR

BMC microbiology 20251103 1

<h4>Background</h4>Understanding bacterial responses to nutrient limitation is critical for developing targeted antimicrobial strategies. Sulfur starvation uniquely induces not only genes responsible for sulfur scavenging but also prominent antioxidant defenses. However, the biological rationale behind the simultaneous induction of antioxidants during sulfur limitation remains largely unexplored. Our study addresses this gap by integrating transcriptomic, proteomic, and targeted metabolomic data ...[more]

PMID: 41184751

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Project description:Among multiple interconnected pathways for L-Lysine (L-Lys) catabolism in pseudomonads, Pseudomonas aeruginosa PAO1 employed the decarboxylase and the transaminase pathways. However, up till now several genes involved in the operation and regulation of these pathways were still missing. Transcriptome analyses coupled with promoter activity measurements and mutant growth phenotype analysis lead us to identify several new members of the L-Lys and D-Lys catabolic pathways and their regulatory elements, including argR to trigger lysine decarboxylation into cadaverine, PauR for the Î³-glutamylation pathway of polyamine catabolism into 5-aminovalerate, gcdR-gcdHG for glutarate utilization, dpkA, amaR-amaAB and PA2035 for D-Lys catabolism, lysR-lysXE for L-Lys efflux, and lysP for L-Lys uptake. Gene expression microarray, including probe preparation, hybridization, fluidics run and chip scan, was performed by Georgia State University DNA/Protein Core Facility. P. aeruginosa PAO1 was grown aerobically in minimal medium P with 350 rpm shaking at 37C, in the presence of 10mM L-glutamate supplemented with 10 mM L-lysine, cadaverine, 5-amino valerate, glutaric acid, D-lysine or 5mM L-pipecolate. Cells were harvested when the optical density at 600 nm reached 0.5~0.6 by centrifugation for 5 minutes at 4C. Total RNA samples were isolated by RNeasy purification kit following instructions of the manufacturer (Qiagen). Reverse transcription for cDNA synthesis, fragmentation by DNase I treatment, cDNA probe labeling and hybridization were performed according to the instructions of GeneChip manufacturer (Affymetrix). Data were processed by Microarray Suite 5.0 software normalizing the absolute expression signal values of all chips to a target intensity of 500. GeneSpring software (Silicon Genetics) was used for expression pattern analysis and comparison. Data collection was carried out using GCOS 1.4 software (Affymetrix). Probe intensity values were normalized to a target value of 500 with normalization factor equal to 1. Data analysis was performed using GeneSpring GX 11 Software (Aglient, Palo Alto,CA). Related Research Papers: 1. Indurthi SM, Chou HT, Lu CD. Molecular Characterization of lysR-lysXE, gcdR-gcdHG, and amaR-amaAB Operons for Lysine Export and Catabolism: A Comprehensive Lysine Catabolic Network in Pseudomonas aeruginosa PAO1. Microbiology. 2015 Submitted [EMID:04803de65c782] 2. Chou HT, Li J, and Lu CD. Functional Characterization of the agtABCD and agtSR Operons for Î³-Aminobutyrate and Î´-Aminovalerate Uptake and Regulation in Pseudomonas aeruginosa PAO1. Curr. Microbiology. 2014 Jan;68(1):59-63. 3. Chou HT, Li JY, Peng YC, Lu CD. Molecular characterization of PauR and its role in control of putrescine and cadaverine catabolism through the Î³-glutamylation pathway in Pseudomonas aeruginosa PAO1. J Bacteriol. 2013 Sep; 195(17):3906-13. 4. Chou HT, Hegazy M, Lu CD. L-lysine Catabolism is Controlled by Arginine/ArgR in Pseudomonas aeruginosa PAO1. J Bacteriol. 2010 Nov;192(22):5874-80

			Action	DRS
	Combined_Proteins_Unfiltered.tsv	Tabular
	PAO1_SS_Manuscript_experimental_protein_list.xlsx	Xlsx
	S2.raw	Raw
	S3.raw	Raw
	S4.raw	Raw