Putative Bifunctional Chorismate Mutase/Prephenate Dehydratase Contributes to the Virulence of Acidovorax citrulli

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

doi:10.3389/fpls.2020.569552

Cited by 14 publications

(30 citation statements)

References 48 publications

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“…The observation here that GqqA has an additional acylase activity may explain some of the earlier observations that have been observed for pheA mutations in other Gram-negative bacteria but that could not be explained by the simple knockout of an amino acid biosynthesis pathway. Among those the recent observations that motility and pathogenicity, which are usually QS-dependent processes in Acidovorax citrulli 35 were altered in the background of a pheA mutant. Further the increased persistence of the opportunistic pathogen Pseudomonas aeruginosa had been reported in a phe mutant and may also hint into a similar direction 36 .…”

Section: Resultsmentioning

confidence: 99%

The Komagataeibacter europaeus GqqA is the prototype of a novel bifunctional N-Acyl-homoserine lactone acylase with prephenate dehydratase activity

Werner¹,

Petersen

Vollstedt

et al. 2021

Preprint

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Previously, we reported the isolation of a quorum quenching protein (QQ), designated GqqA, from Komagataeibacter europaeus CECT 8546 that is highly homologous to prephenate dehydratases (PDT) 1. GqqA strongly interfered with N-acyl-homoserine lactone (AHL) quorum sensing signals from Gram-negative bacteria and affected biofilm formation in its native host strain Komagataeibacter europaeus. Here we present and discuss data identifying GqqA as a novel acylase. ESI-MS-MS data showed unambiguously that GqqA hydrolyzes the amide bond of the acyl side-chain of AHL molecules, but not the lactone ring. Consistent with this observation the protein sequence does not carry a conserved Zn2+ binding motif, known to be essential for metal-dependent lactonases, but in fact harboring the typical periplasmatic binding protein domain (PBP domain), acting as catalytic domain. We report structural details for the native structure at 2.5 Å resolution and for a truncated GqqA structure at 1.7 Å. The structures obtained highlight that GqqA acts as a dimer and complementary docking studies indicate that the lactone ring of the substrate binds within a cleft of the PBP domain and interacts with polar residues Y16, S17 and T174. The biochemical and phylogenetic analyses imply that GqqA represents the first member of a novel type of QQ family enzymes.

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Section: Resultsmentioning

confidence: 99%

The Komagataeibacter europaeus GqqA is the prototype of a novel bifunctional N-Acyl-homoserine lactone acylase with prephenate dehydratase activity

Werner¹,

Petersen

Vollstedt

et al. 2021

Preprint

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“…The planted seeds were then grown in a growth chamber and checked for disease severity for 7 days. Disease severity was evaluated using a 0-2 scale (0, no symptoms; 1, water-soaked region; 2, seedling wilt) as described previously (Kim et al, 2020;Song et al, 2020). A disease index was then calculated as follows:…”

Section: Generation Of Acδglpdac and Its Complementary Strainmentioning

confidence: 99%

“…It is clear that GlpdAc is involved in virulence. In addition, it has been shown that one protein related to biochemical pathways can produce pleiotropic effects in bacteria, in- cluding the alteration of virulence and other physiological mechanisms (Felgner et al, 2016;Kim et al, 2020). Therefore, we carried out comparative proteomics to postulate biological and cellular mechanisms related to GlpdAc using the wild-type and AcΔglpdAc strains.…”

Section: Comparative Proteomics Analysis In Ac and Acδglpdacmentioning

confidence: 99%

“…Additionally, research focusing on the type III secretion system is actively ongoing (Jiménez-Guerrero et al, 2019). Recently, Kim et al (2020) reported that CmpAc, which is a bifunctional chorismate mutase and prephenate dehydratase, is involved in virulence and other mechanisms in Ac. However, other genes and mechanisms that are related to virulence in Ac have not been definitively identified.…”

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confidence: 99%

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Proteomic and Phenotypic Analyses of a Putative Glycerol-3-Phosphate Dehydrogenase Required for Virulence in Acidovorax citrulli

et al. 2021

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Acidovorax citrulli (Ac) is the causal agent of bacterial fruit blotch (BFB) in watermelon, a disease that poses a serious threat to watermelon production. Because of the lack of resistant cultivars against BFB, virulence factors or mechanisms need to be elucidated to control the disease. Glycerol-3-phosphate dehydrogenase is the enzyme involved in glycerol production from glucose during glycolysis. In this study, we report the functions of a putative glycerol-3-phosphate dehydrogenase in Ac (GlpdAc) using comparative proteomic analysis and phenotypic observation. A glpdAc knockout mutant, AcΔglpdAc(EV), lost virulence against watermelon in two pathogenicity tests. The putative 3D structure and amino acid sequence of GlpdAc showed high similarity with glycerol-3-phosphate dehydrogenases from other bacteria. Comparative proteomic analysis revealed that many proteins related to various metabolic pathways, including carbohydrate metabolism, were affected by GlpdAc. Although AcΔglpdAc(EV) could not use glucose as a sole carbon source, it showed growth in the presence of glycerol, indicating that GlpdAc is involved in glycolysis. AcΔglpdAc(EV) also displayed higher cellto-cell aggregation than the wild-type bacteria, and tolerance to osmotic stress and ciprofloxacin was reduced and enhanced in the mutant, respectively. These results indicate that GlpdAc is involved in glycerol metabolism and other mechanisms, including virulence, demonstrating that the protein has pleiotropic effects. Our study expands the understanding of the functions of proteins associated with virulence in Ac.

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“…Moreover, the type IV pili-mediated motility and quorum-sensing are required for its virulence and seed-transmission ( Bahar et al, 2009 ; Johnson and Walcott, 2013 ). Recently, two proteins, bifunctional chorismate mutase/prephenate dehydratase (CmpAc) and glycerol-3-phosphate dehydrogenase (GlpdAc), have been identified in Ac ( Kim et al, 2020b , 2021 ). Both proteins are involved in its virulence as well as in other mechanisms, such as twitching motility, tolerance against external stresses, or growth in diverse carbon sources.…”

mentioning

confidence: 99%

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

2021

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Acidovorax citrulli (Ac) is the causative agent of bacterial fruit blotch disease in watermelon. Since resistant cultivars have not yet been developed, the virulence factors/mechanisms of Ac need to be characterized. This study reports the functions of a putative pyridoxal phosphate-dependent aminotransferase (PpdaAc) that transfers amino groups to its substrates and uses pyridoxal phosphate as a coenzyme. It was observed that a ppdaAc knockout mutant had a significantly reduced virulence in watermelon when introduced via germinated-seed inoculation as well as leaf infiltration. Comparative proteomic analysis predicted the cellular mechanisms related to PpdaAc. Apart from causing virulence, the PpdaAc may have significant roles in energy production, cell membrane, motility, chemotaxis, post-translational modifications, and iron-related mechanisms. Therefore, it is postulated that PpdaAc may possess pleiotropic effects. These results provide new insights into the functions of a previously unidentified PpdaAc in Ac.

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Putative Bifunctional Chorismate Mutase/Prephenate Dehydratase Contributes to the Virulence of Acidovorax citrulli

Cited by 14 publications

References 48 publications

The Komagataeibacter europaeus GqqA is the prototype of a novel bifunctional N-Acyl-homoserine lactone acylase with prephenate dehydratase activity

The Komagataeibacter europaeus GqqA is the prototype of a novel bifunctional N-Acyl-homoserine lactone acylase with prephenate dehydratase activity

Proteomic and Phenotypic Analyses of a Putative Glycerol-3-Phosphate Dehydrogenase Required for Virulence in Acidovorax citrulli

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

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