The<i>Erwinia chrysanthemi</i>3937 PhoQ Sensor Kinase Regulates Several Virulence Determinants

Balakrishnan, Vallath; Babujee, Lavanya; Liu, Hui; Hedley, Pete E.; Fujikawa, Takashi; Birch, Paul R. J.; Toth, Ian K.; Tsuyumu, Shinji

doi:10.1128/jb.188.8.3088-3098.2006

Cited by 49 publications

(65 citation statements)

References 61 publications

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“…For each array, two total RNA samples (3 g each) were indirectly labeled by reverse transcription using an 11-mer oligonucleotide mixture in the presence of amino-allyl dUTP, as described (30). Labeled cDNA samples were then coupled with Cy3 or Cy5 dye esters (GE Healthcare) and purified (30). Cy-labeled wild type and mutant cDNAs were co-hybridized to Agilent Technologies, Inc., custom ECA_11K Genome Arrays according to the manufacturer's instructions and according to Ref.…”

Section: Methodsmentioning

confidence: 99%

“…Cy-labeled wild type and mutant cDNAs were co-hybridized to Agilent Technologies, Inc., custom ECA_11K Genome Arrays according to the manufacturer's instructions and according to Ref. 30. The probes (60-mers) include all 4521 predicted protein coding genes and 23 Lucidea Universal ScoreCard Controls (GE Healthcare), with each probe printed twice.…”

Section: Methodsmentioning

confidence: 99%

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DsbA Plays a Critical and Multifaceted Role in the Production of Secreted Virulence Factors by the Phytopathogen Erwinia carotovora subsp. atroseptica

Coulthurst

Lilley

Hedley³

et al. 2008

Journal of Biological Chemistry

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Erwinia carotovora subsp. atroseptica is an enterobacterial phytopathogen causing economically significant soft rot disease. Pathogenesis is mediated by multiple secreted virulence factors, many of which are secreted by the type II (Out) secretion system. DsbA catalyzes the introduction of disulfide bonds into periplasmic and secreted proteins. In this study, the extracellular proteome (secretome) of wild type E. carotovora subsp. atroseptica SCRI1043, and dsbA and out mutants, was analyzed by spectral counting mass spectrometry. This revealed that dsbA inactivation had a huge impact on the secretome and identified diverse DsbA-and Out-dependent secreted proteins, representing known, predicted, and novel candidate virulence factors. Further characterization of the dsbA mutant showed that secreted enzyme activities, motility, production of the quorumsensing signal, and virulence were absent or substantially reduced. The impact of DsbA on secreted virulence factor production was mediated at multiple levels, including impacting on the Out secretion system and the virulence gene regulatory network. Transcriptome analyses revealed that the abundance of a broad, but defined, set of transcripts, including many virulence factors, was altered in the dsbA mutant, identifying a new virulence regulon responsive to extracytoplasmic conditions. In conclusion, DsbA plays a crucial, multifaceted role in the pathogenesis of E. carotovora subsp. atroseptica.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

DsbA Plays a Critical and Multifaceted Role in the Production of Secreted Virulence Factors by the Phytopathogen Erwinia carotovora subsp. atroseptica

Coulthurst

Lilley

Hedley³

et al. 2008

Journal of Biological Chemistry

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“…Indeed, at least six of the genes present in this cluster have been described as being induced and/or implicated in responses to various stress conditions. They include trmD, encoding a tRNA (guanine-n1) methyltransferase (55); rpsU, encoding the 30S ribosomal protein s21 (35); cspA, encoding a conserved transcription and translation enhancer (39); and acnA, encoding an aconitate hydratase (26,56). Perhaps most interesting in this cluster is ompR, encoding the response regulator from the EnvZ/OmpR two-component system, identi- Table S2 in the supplemental material, and the conservation score is given in Table S3 in the supplemental material.…”

Section: Discussionmentioning

confidence: 99%

The BatR/BatS Two-Component Regulatory System Controls the Adaptive Response ofBartonella henselaeduring Human Endothelial Cell Infection

et al. 2010

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Here, we report the first comprehensive study of Bartonella henselae gene expression during infection of human endothelial cells. Expression of the main cluster of upregulated genes, comprising the VirB type IV secretion system and its secreted protein substrates, is shown to be under the positive control of the transcriptional regulator BatR. We demonstrate binding of BatR to the promoters of the virB operon and a substrate-encoding gene and provide biochemical evidence that BatR and BatS constitute a functional twocomponent regulatory system. Moreover, in contrast to the acid-inducible (pH 5.5) homologs ChvG/ChvI of Agrobacterium tumefaciens, BatR/BatS are optimally activated at the physiological pH of blood (pH 7.4). By conservation analysis of the BatR regulon, we show that BatR/BatS are uniquely adapted to upregulate a genus-specific virulence regulon during hemotropic infection in mammals. Thus, we propose that BatR/BatS two-component system homologs represent vertically inherited pH sensors that control the expression of horizontally transmitted gene sets critical for the diverse host-associated life styles of the alphaproteobacteria.

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“…The nine genes selected for analysis encode proteins involved in different metabolic activities. rplI and rplQ (30), glnA, rho, recA, gyrA, gyrB, and pro1A (54), and 16S rRNA are housekeeping genes that have been used as reference genes in similar studies of various bacteria (51,55). Of the nine genes tested, only the 16S rRNA gene was stably expressed at the two temperatures.…”

Section: Discussionmentioning

confidence: 99%

Identification of Cold-Temperature-Regulated Genes in Flavobacterium psychrophilum

Hesami

Metcalf

Lumsden

et al. 2011

Appl Environ Microbiol

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Flavobacterium psychrophilum is the etiological agent of bacterial coldwater disease (BCWD) and rainbow trout fry syndrome (RTFS). It causes disease primarily in fresh water-reared salmonids, but other fish species can also be affected. A diverse array of clinical conditions is associated with BCWD, including tail rot (peduncle disease), necrotic myositis, and cephalic osteochondritis. Degradation of connective and muscular tissues by extracellular proteases is common to all of these presentations. There are no effective vaccines to prevent BCWD or RTFS, and antibiotics are often used to prevent and control disease. To identify virulence factors that might permit development of an efficacious vaccine, cDNA suppression subtractive hybridization (SSH) was used to identify cold-regulated genes in a virulent strain of F. psychrophilum. Genes predicted to encode a two-component system sensor histidine kinase (LytS), an ATP-dependent RNA helicase, a multidrug ABC transporter permease/ATPase, an outer membrane protein/protective antigen OMA87, an M43 cytophagalysin zinc-dependent metalloprotease, a hypothetical protein, and four housekeeping genes were upregulated at 8°C versus the level of expression at 20°C. Because no F. psychrophilum gene was known to be suitable as an internal standard in reverse transcription-quantitative real-time PCR (RT-qPCR) experiments, the expression stability of nine commonly used reference genes was evaluated at 8°C and 20°C. Expression of the 16S rRNA was equivalent at both temperatures, and this gene was used in RT-qPCR experiments to verify the SSH findings. With the exception of the ATCC 49513 strain, similar patterns of gene expression were obtained with 11 other representative strains of F. psychrophilum.Bacterial cold water disease (BCWD), caused by Flavobacterium psychrophilum, occurs at low water temperatures and can cause economic losses in the aquaculture industry as a result of direct mortality or vertebral deformities that decrease the market value of fish that survive the infection (31, 35). A number of putative virulence factors of this bacterium have been identified, including extracellular proteases involved in degradation of extracellular matrix components such as elastin, fibrinogen, type IV collagen, actin, and myosin (9,37,47,48). The best studied of these, psychrophilic metalloproteases Fpp1 and Fpp2, are thought to be involved in destruction of host tissues; however, their roles have not yet been demonstrated in vivo (47,48). In a recent study, Sudheesh et al. (53) used two-dimensional acrylamide gel electrophoresis and Western blot analysis to compare a virulent strain and a nonvirulent strain of F. psychrophilum and found a thermolysin that was unique to the virulent strain. A role for the iron uptake-associated gene exbD2 has also been demonstrated. Strains lacking this gene have decreased virulence and confer a high level of protection in rainbow trout fry when they are used for vaccination (4). Other putative virulence factors of F. psychrophilum include a lipo...

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TheErwinia chrysanthemi3937 PhoQ Sensor Kinase Regulates Several Virulence Determinants

Cited by 49 publications

References 61 publications

DsbA Plays a Critical and Multifaceted Role in the Production of Secreted Virulence Factors by the Phytopathogen Erwinia carotovora subsp. atroseptica

DsbA Plays a Critical and Multifaceted Role in the Production of Secreted Virulence Factors by the Phytopathogen Erwinia carotovora subsp. atroseptica

The BatR/BatS Two-Component Regulatory System Controls the Adaptive Response ofBartonella henselaeduring Human Endothelial Cell Infection

Identification of Cold-Temperature-Regulated Genes in Flavobacterium psychrophilum

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