Comparative Proteomic Analysis for a Putative Pyridoxal Phosphate-Dependent Aminotransferase Required for Virulence in Acidovorax citrulli

Lee, Jong-Chan; Heo, Lynn; Han, Sang-Wook

doi:10.5423/ppj.nt.09.2021.0139

Cited by 5 publications

(9 citation statements)

References 39 publications

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“…Supplementary Table 1 organizes all plasmids and bacterial strains used in this study. AcΔyppAc was selected via screening the Tn5 insertional mutant library using a previously reported method ( Kim et al, 2020 ; Lee et al, 2021 ). A germinated seed inoculation was used to select virulence-deficient mutants.…”

Section: Methodsmentioning

confidence: 99%

“…Another study verified that type IV pili influence twitching motility, surface adhesion, and virulence ( Bahar et al, 2009 ). Quorum sensing genes, ferric uptake regulators, and a putative glycerol-3-phosphate dehydrogenase are required for Ac virulence ( Johnson and Walcott, 2013 ; Lee et al, 2021 ; Liu et al, 2019 ). However, most studies only focus on the group I strains.…”

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confidence: 99%

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Proteomic and Phenotypic Analyses of a Putative YggS Family Pyridoxal Phosphate-Dependent Enzyme in Acidovorax citrulli

et al. 2023

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Acidovorax citrulli (Ac) is a phytopathogenic bacterium that causes bacterial fruit blotch (BFB) in cucurbit crops, including watermelon. However, there are no effective methods to control this disease. YggS family pyridoxal phosphate-dependent enzyme acts as a coenzyme in all transamination reactions, but its function in Ac is poorly understood. Therefore, this study uses proteomic and phenotypic analyses to characterize the functions. The Ac strain lacking the YggS family pyridoxal phosphate-dependent enzyme, AcΔyppAc(EV), virulence was wholly eradicated in geminated seed inoculation and leaf infiltration. AcΔyppAc(EV) propagation was inhibited when exposed to L-homoserine but not pyridoxine. Wild-type and mutant growth were comparable in the liquid media but not in the solid media in the minimal condition. The comparative proteomic analysis revealed that YppAc is primarily involved in cell motility and wall/membrane/envelop biogenesis. In addition, AcΔyppAc(EV) reduced biofilm formation and twitching halo production, indicating that YppAc is involved in various cellular mechanisms and possesses pleiotropic effects. Therefore, this identified protein is a potential target for developing an efficient anti-virulence reagent to control BFB.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

Proteomic and Phenotypic Analyses of a Putative YggS Family Pyridoxal Phosphate-Dependent Enzyme in Acidovorax citrulli

et al. 2023

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“…Five independent experiments with ten biological replicates were conducted per strain. The leaf infiltration experiments were conducted as previously reported ( Lee et al., 2021 ). Briefly, the watermelon seeds were grown in a growth chamber until they reached the four true leaves stage.…”

Section: Methodsmentioning

confidence: 99%

“…Additionally, quorum sensing, a cell-to-cell communication system in bacteria that can regulate gene expression, is also a well-known virulence mechanism in gram-negative pathogenic bacteria including Ac (De Kievit and Iglewski, 2000;Wang et al, 2016). Previous studies have also reported that a ferric uptake regulator, yggS family pyridoxal 5'-phosphate binding protein, a bifunctional chorismate mutase/prephenate dehydratase, and a pyridoxal phosphate-dependent aminotransferase contribute to virulence in Ac (Liu et al, 2019;Kim et al, 2020;Lee et al, 2021;Wang et al, 2022). Although several studies have characterized Ac virulence, additional efforts are still needed, as many virulence-related genes in Ac have not been studied yet.…”

Section: Introductionmentioning

confidence: 99%

A putative 2,3-bisphosphoglycerate-dependent phosphoglycerate mutase is involved in the virulence, carbohydrate metabolism, biofilm formation, twitching halo, and osmotic tolerance in Acidovorax citrulli

Lee

Cho

et al. 2022

Front. Plant Sci.

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Acidovorax citrulli (Ac) is a gram-negative bacterium that causes bacterial fruit blotch (BFB) disease in cucurbit crops including watermelon. However, despite the great economic losses caused by this disease worldwide, Ac-resistant watermelon cultivars have not been developed. Therefore, characterizing the virulence factors/mechanisms of Ac would enable the development of effective control strategies against BFB disease. The 2,3-bisphosphoglycerate-dependent phosphoglycerate mutase (BdpM) is known to participate in the glycolysis and gluconeogenesis pathways. However, the roles of the protein have not been characterized in Ac. To elucidate the functions of BdpmAc (Bdpm in Ac), comparative proteomic analysis and diverse phenotypic assays were conducted using a bdpmAc knockout mutant (bdpmAc:Tn) and a wild-type strain. The virulence of the mutant to watermelon was remarkably reduced in both germinated seed inoculation and leaf infiltration assays. Moreover, the mutant could not grow with fructose or pyruvate as a sole carbon source. However, the growth of the mutant was restored to levels similar to those of the wild-type strain in the presence of both fructose and pyruvate. Comparative proteomic analyses revealed that diverse proteins involved in motility and wall/membrane/envelop biogenesis were differentially abundant. Furthermore, the mutant exhibited decreased biofilm formation and twitching halo size. Interestingly, the mutant exhibited a higher tolerance against osmotic stress. Overall, our findings suggest that BdpmAc affects the virulence, glycolysis/gluconeogenesis, biofilm formation, twitching halo size, and osmotic tolerance of Ac, suggesting that this protein has pleiotropic properties. Collectively, our findings provide fundamental insights into the functions of a previously uncharacterized phosphoglycerate mutase in Ac.

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“…The complete genome sequence of this strain was recently reported (Kang et al, 2021). For comparative proteomic analysis, we used a label-free shotgun proteomic technique, following a previously established protocol (Lee et al, 2021). The strain was cultured in three different liquid media: the Luria Bertani medium (LB; 10 g of tryptone, 5 g of yeast extract, and 10 g of NaCl in 1 l), an hrp inducing medium (HMM; 1 g of (NH 4 ) 2 SO 4 , 0.246 g of MgCl 2 -6H 2 O, 0.1 g of NaCl, 8.708 g of K 2 HPO 4 , and 6.804 g of KH 2 PO 4 in 1 l), and an amylovoran inducing medium (MBMA; 3 g of KH 2 PO 4 , 7 g of K 2 HPO 4 , 1 g of (NH 4 ) 2 SO 4 , 2 ml of glycerol, 0.5 g of citric acid, and 0.03 g of MgSO 4 , and 10 g of sorbitol in 1 l).…”

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confidence: 99%

Comparing Protein Expression in Erwinia amylovora Strain TS3128 Cultured under Three Sets of Environmental Conditions

et al. 2022

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Erwinia amylovora, the causal agent of fire-blight disease in apple and pear trees, was first isolated in South Korea in 2015. Although numerous studies, including omics analyses, have been conducted on other strains of E. amylovora, studies on South Korean isolates remain limited. In this study, we conducted a comparative proteomic analysis of the strain TS3128, cultured in three media representing different growth conditions. Proteins related to virulence, type III secretion system, and amylovoran production, were more abundant under minimal conditions than in rich conditions. Additionally, various proteins associated with energy production, carbohydrate metabolism, cell wall/membrane/envelope biogenesis, and ion uptake were identified under minimal conditions. The strain TS3128 expresses these proteins to survive in harsh environments. These findings contribute to understanding the cellular mechanisms driving its adaptations to different environmental conditions and provide proteome profiles as reference for future studies on the virulence and adaptation mechanisms of South Korean strains.

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Comparative Proteomic Analysis for a Putative Pyridoxal Phosphate-Dependent Aminotransferase Required for Virulence in Acidovorax citrulli

Cited by 5 publications

References 39 publications

Proteomic and Phenotypic Analyses of a Putative YggS Family Pyridoxal Phosphate-Dependent Enzyme in Acidovorax citrulli

Proteomic and Phenotypic Analyses of a Putative YggS Family Pyridoxal Phosphate-Dependent Enzyme in Acidovorax citrulli

A putative 2,3-bisphosphoglycerate-dependent phosphoglycerate mutase is involved in the virulence, carbohydrate metabolism, biofilm formation, twitching halo, and osmotic tolerance in Acidovorax citrulli

Comparing Protein Expression in Erwinia amylovora Strain TS3128 Cultured under Three Sets of Environmental Conditions

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