HrpG regulates type II secretory proteins in Xanthomonas axonopodis pv. citri

Yamazaki, Akihiro; Hirata, Haruyuki; Tsuyumu, Shinji

doi:10.1007/s10327-008-0075-7

Cited by 39 publications

(57 citation statements)

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“…Even though the two-component system has been suggested to play crucial roles in multiple cellular functions of X. citri subsp. citri, only a few have been studied for their contributions to virulence, such as HrpG (46). In this study, we present evidence that the two-component system ColR/ColS plays important roles in the virulence, growth in planta, biofilm formation, lipopolysaccharide (LPS) production, catalase activity, and environmental stress resistance of the citrus canker pathogen X. citri subsp.…”

mentioning

confidence: 91%

The ColR/ColS Two-Component System Plays Multiple Roles in the Pathogenicity of the Citrus Canker Pathogen Xanthomonas citri subsp. citri

Yan

Wang

2011

J Bacteriol

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Bacterial citrus canker disease, which is caused by Xanthomonas citri subsp. citri, is one of the most devastating diseases of citrus plants. In this study, we characterized the role of the two-component regulatory system ColR/ColS in the pathogenicity of X. citri subsp. citri. colS mutants (256A10 and 421E7), colR mutants (386C6 and 417E10), and a colR colS double mutant (306DSR) all lost pathogenicity and produced no symptoms on grapefruit leaves inoculated by either pressure infiltration or the spray method. The pathogenicity defect of the colS, colR, and colR colS mutants could be complemented using the wild-type colS, colR, and colR colS genes, respectively. Mutation of colS or colR significantly reduced X. citri subsp. citri growth in planta. The ColR/ColS system also played important roles in bacterial biofilm formation in glass tubes and on leaf surfaces, lipopolysaccharide (LPS) production, catalase activity, and tolerance of environmental stress, including phenol, copper, and hydrogen peroxide. Furthermore, quantitative reverse transcription-PCR assays demonstrated that the ColR/ColS system positively regulated the expression of important virulence genes, including hrpD6, hpaF, the O-antigen LPS synthesis gene rfbC, and the catalase gene katE. Overall, our data indicate that the two-component regulatory system ColR/ColS is critical for X. citri subsp. citri virulence, growth in planta, biofilm formation, catalase activity, LPS production, and resistance to environmental stress. Xanthomonas

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confidence: 91%

The ColR/ColS Two-Component System Plays Multiple Roles in the Pathogenicity of the Citrus Canker Pathogen Xanthomonas citri subsp. citri

Yan

Wang

2011

J Bacteriol

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“…The regulation of hrp genes might be influenced by plant-inducing factors that occur owing to the presence of a plant-inducible promoter known as PIP box [TTCGC (N 15 ) TTCGC], as well as the nutritional conditions of the medium (Leach and White, 1996;Da Silva et al, 2002). Yamazaki et al (2008) have suggested the presence of an imperfect PIP box for the CPXAC gene. By contrast, pathogenicity genes hrpX and hrpG are responsible for the secretion of proteins in type III and some type II systems (Sandkvist, 2001;Furutani et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

“…The secretion products from Xac have been described by Yamazaki et al (2008), who reported the presence of an extracellular cysteine peptidase secreted by Xac (strain NA-1) cultivated in medium that induced hrp and hypersensitivity genes. The N-terminal region of this secreted cysteine peptidase was sequenced and generated AVHGMGLKPSSLMQPV, which is identical to the sequence from HIS CPXAC (after the 25th residue).…”

Section: Discussionmentioning

confidence: 99%

“…The N-terminal region of this secreted cysteine peptidase was sequenced and generated AVHGMGLKPSSLMQPV, which is identical to the sequence from HIS CPXAC (after the 25th residue). This match was important for the identification and characterization of the target protein because it revealed the iden-tity of the N-terminal sequence of the authentic secretion product from Xanthomonas with CPXAC, suggesting the enzyme identified by Yamazaki et al (2008) to be CPXAC.…”

Section: Discussionmentioning

confidence: 99%

“…Particular attention has been paid to extracellular peptidases owing to their possible role in bacterial pathogenesis and their usefulness as molecular targets for understanding and controlling diseases. Yamazaki et al (2008) have identified several proteins secreted from Xanthomonas pv. citri NA-1 cultivated under conditions conducive for the induction of hrp and hypersensitivity genes, which may be related to pathogenicity and the infection process.…”

Section: Introductionmentioning

confidence: 99%

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Recombinant expression and characterization of a cysteine peptidase from Xanthomonas citri subsp citri

Soares-Costa¹,

Silveira²,

Novo³

et al. 2012

Genet. Mol. Res.

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ABSTRACT. Xanthomonas citri subsp citri (Xac) is the bacterium responsible for citrus canker disease in citrus plants. The aim of this study was to describe the recombinant expression, purification, and characterization of a cysteine peptidase from Xac strain 306, which is a candidate for involvement in the pathogenicity of this bacterium. The gene was cloned and expressed in Pichia pastoris, and the cysteine peptidase was successfully expressed, secreted, and purified using affinity chromatography with a yield of approximately 10 mg/L. A polyclonal antibody produced against cysteine peptidase from X. citri subsp citri fused with HIS tag ( HIS CPXAC) recognized the purified recombinant cysteine peptidase HIS CPXAC, confirming the correct production of this protein in P. pastoris. The same antibody detected the protein in the culture supernatant of Xac grown in pathogenicityinducing medium. Kinetic analysis revealed that HIS CPXAC hydrolyzed the carbobenzoxy-Leu-Arg-7-amido-4-methylcoumarin substrate with a catalytic efficiency (k cat /K m ) of 47 μM -1 •s -1 . The purified HIS CPXAC displayed maximal catalytic activity at pH 5.5 and 30°C. The recombinant enzyme was inhibited by the specific cysteine peptidase inhibitor E-64, as well as by the recombinant cysteine peptidase inhibitors CaneCPI-1, CaneCPI-2, CaneCPI-3, and CaneCPI-4, with K i values of 1.214, 84.64, 0.09, 0.09, and 0.012 nM, respectively. Finally, the N-terminal sequencing of the purified protein enabled the identification of the first 5 amino acid residues (AVHGM) immediately after the putative signal peptide, thereby enabling the identification of the cleavage point and corroborating previous studies that have identified this sequence in a secreted protein from Xanthomonas spp.

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Bacterial adaptation to life in association with plants – A proteomic perspective from culture to in situ conditions

2011

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Diverse bacterial taxa that live in association with plants affect plant health and development. This is most evident for those bacteria that undergo a symbiotic association with plants or infect the plants as pathogens. Proteome analyses have contributed significantly toward a deeper understanding of the molecular mechanisms underlying the development of these associations. They were applied to obtain a general overview of the protein composition of these bacteria, but more so to study effects of plant signaling molecules on the cytosolic proteome composition or metabolic adaptations upon plant colonization. Proteomic analyses are particularly useful for the identification of secreted proteins, which are indispensable to manipulate a host plant. Recent advances in the field of proteome analyses have initiated a new research area, the analysis of more complex microbial communities. Such studies are just at their beginning but hold great potential for the future to elucidate not only the interactions between bacteria and their host plants, but also of bacteria-bacteria interactions between different bacterial taxa when living in association with plants. These include not only the symbiotic and pathogenic bacteria, but also the commensal bacteria that are consistently found in association with plants and whose functions remain currently largely uncovered.

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HrpG regulates type II secretory proteins in Xanthomonas axonopodis pv. citri

Cited by 39 publications

References 42 publications

The ColR/ColS Two-Component System Plays Multiple Roles in the Pathogenicity of the Citrus Canker Pathogen Xanthomonas citri subsp. citri

The ColR/ColS Two-Component System Plays Multiple Roles in the Pathogenicity of the Citrus Canker Pathogen Xanthomonas citri subsp. citri

Recombinant expression and characterization of a cysteine peptidase from Xanthomonas citri subsp citri

Bacterial adaptation to life in association with plants – A proteomic perspective from culture to in situ conditions

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