Characterization of two distinct phospholipase C enzymes from Burkholderia pseudomallei

Korbsrisate, Sunee; Tomaras, Andrew P.; Damnin, Suwat; Ckumdee, Jutturong; Srinon, Varintip; Lengwehasatit, I.; Vasil, Michael L.; Suparak, Supaporn

doi:10.1099/mic.0.2006/003004-0

Cited by 45 publications

(44 citation statements)

References 35 publications

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“…3B). Korbsrisate et al demonstrated that B. pseudomallei PlcN1 and PlcN2 exhibit extracellular PLC activity (40). We found that PlcN1 and PlcN2 were secreted in a T2SS-dependent manner (Table 2), and it is likely that they both contribute to the PLC activity present in the supernatants of MSHR668 and 668 ⌬gspD/gspD (Fig.…”

Section: Construction and Initial Characterization Of B Pseudomalleimentioning

confidence: 52%

“…Thus, it appears that the B. mallei T2SS is functional and that it exports TssM, but this will have to be confirmed in future studies. Finally, the B. mallei genome has undergone numerous deletion and rearrangement events during the host adaptation process, and two of the genes that were "lost" are mprA and plcN2 (39,40,57). We propose that the loss of these genes might be responsible for the protease and PLC secretion deficiencies of B. mallei rather than the putative mutations in the B. mallei T2SS gene cluster (57).…”

Section: Discussionmentioning

confidence: 95%

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Proteomic Analysis of the Burkholderia pseudomallei Type II Secretome Reveals Hydrolytic Enzymes, Novel Proteins, and the Deubiquitinase TssM

2014

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bBurkholderia pseudomallei, the etiologic agent of melioidosis, is an opportunistic pathogen that harbors a wide array of secretion systems, including a type II secretion system (T2SS), three type III secretion systems (T3SS), and six type VI secretion systems (T6SS). The proteins exported by these systems provide B. pseudomallei with a growth advantage in vitro and in vivo, but relatively little is known about the full repertoire of exoproducts associated with each system. In this study, we constructed deletion mutations in gspD and gspE, T2SS genes encoding an outer membrane secretin and a cytoplasmic ATPase, respectively. The secretion profiles of B. pseudomallei MSHR668 and its T2SS mutants were noticeably different when analyzed by SDS-PAGE. We utilized liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify proteins present in the supernatants of B. pseudomallei MSHR668 and B. pseudomallei ⌬gspD grown in rich and minimal media. The MSHR668 supernatants contained 48 proteins that were either absent or substantially reduced in the supernatants of ⌬gspD strains. Many of these proteins were putative hydrolytic enzymes, including 12 proteases, two phospholipases, and a chitinase. Biochemical assays validated the LC-MS/MS results and demonstrated that the export of protease, phospholipase C, and chitinase activities is T2SS dependent. Previous studies had failed to identify the mechanism of secretion of TssM, a deubiquitinase that plays an integral role in regulating the innate immune response. Here we present evidence that TssM harbors an atypical signal sequence and that its secretion is mediated by the T2SS. This study provides the first in-depth characterization of the B. pseudomallei T2SS secretome.

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Section: Construction and Initial Characterization Of B Pseudomalleimentioning

confidence: 52%

Section: Discussionmentioning

confidence: 95%

Proteomic Analysis of the Burkholderia pseudomallei Type II Secretome Reveals Hydrolytic Enzymes, Novel Proteins, and the Deubiquitinase TssM

2014

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“…The redundancy of these phospholipid-degrading enzymes suggests that there is some functional specificity. To this end, studies of the Plc-1 and Plc-2 phospholipases C from B. pseudomallei (orthologues of which are present in the J2315 genome; BCAL1046 and BCAM2429, respectively) have demonstrated that although they both hydrolyze phospholipid phosphatidylcholine and sphingomyelin, they exhibit marked differences in their cytotoxicity (64).…”

Section: General Features Of the J2315 Genomementioning

confidence: 99%

The Genome of Burkholderia cenocepacia J2315, an Epidemic Pathogen of Cystic Fibrosis Patients

et al. 2009

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Bacterial infections of the lungs of cystic fibrosis (CF) patients cause major complications in the treatment of this common genetic disease.Burkholderia cenocepacia infection is particularly problematic since this organism has high levels of antibiotic resistance, making it difficult to eradicate; the resulting chronic infections are associated with severe declines in lung function and increased mortality rates. B. cenocepacia strain J2315 was isolated from a CF patient and is a member of the epidemic ET12 lineage that originated in Canada or the United Kingdom and spread to Europe. The 8.06-Mb genome of this highly transmissible pathogen comprises three circular chromosomes and a plasmid and encodes a broad array of functions typical of this metabolically versatile genus, as well as numerous virulence and drug resistance functions. Although B. cenocepacia strains can be isolated from soil and can be pathogenic to both plants and man, J2315 is representative of a lineage of B. cenocepacia rarely isolated from the environment and which spreads between CF patients. Comparative analysis revealed that ca. 21% of the genome is unique in comparison to other strains of B. cenocepacia, highlighting the genomic plasticity of this species. Pseudogenes in virulence determinants suggest that the pathogenic response of J2315 may have been recently selected to promote persistence in the CF lung. The J2315 genome contains evidence that its unique and highly adapted genetic content has played a significant role in its success as an epidemic CF pathogen.

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“…A different two-component regulatory system known as irlRS has been characterized previously in B. pseudomallei, and its mutant was deficient in the ability to invade a human type II pneumocyte cell line, but it is not a virulence determinant in the infant diabetic rat and Syrian hamster models of acute B. pseudomallei infection (14). In addition, a different phospholipase C gene encoding a phosphatidylcholine-hydrolyzing phospholipase C (PC-PLC) has also been characterized previously in B. pseudomallei, and its product was recognized by human immunoglobulin M antibodies (18). However, there has been no report of this gene's involvement in bacterial virulence.…”

Section: Vol 74 2006mentioning

confidence: 99%

Genome-Wide Expression Analysis of Burkholderia pseudomallei Infection ina Hamster Model of Acute Melioidosis

Tuanyok

Tom

Dunbar³

et al. 2006

Infect Immun

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Burkholderia pseudomallei is the causative agent of melioidosis and represents a potential bioterrorism threat. In the current studies we have examined gene expression in B. pseudomallei in an animal model of acute melioidosis using whole-genome microarrays. Gene expression profiles were generated by comparing transcriptional levels of B. pseudomallei-expressed genes in infected hamster organs including liver, lung, and spleen following intraperitoneal and intranasal routes of infection to those from bacteria grown in vitro. Differentially expressed genes were similar in infected livers irrespective of the route of infection. Reduced expression of a number of housekeeping genes suggested a lower bacterial growth rate during infection. Energy production during growth in vivo involved specific biochemical pathways such as isomerization of 3-phosphoglycerate, catabolism of D-glucosamine and inositol, and biosynthesis of particular amino acids. In addition, the induction of genes known to be involved in oxidative phosphorylation including ubiquinol oxidase, ferredoxin oxidoreductase, and formate dehydrogenase enzymes suggested the use of alternative pathways for energy production, while the expression of genes coding for ATPsynthase and NADH-dehydrogenase enzymes was reduced. Our studies have identified differentially expressed genes which include potential virulence genes such as those for a putative phospholipase C and a putative two-component regulatory system, and they have also provided a better understanding of bacterial metabolism in response to the host environment during acute melioidosis.

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Characterization of two distinct phospholipase C enzymes from Burkholderia pseudomallei

Cited by 45 publications

References 35 publications

Proteomic Analysis of the Burkholderia pseudomallei Type II Secretome Reveals Hydrolytic Enzymes, Novel Proteins, and the Deubiquitinase TssM

Proteomic Analysis of the Burkholderia pseudomallei Type II Secretome Reveals Hydrolytic Enzymes, Novel Proteins, and the Deubiquitinase TssM

The Genome of Burkholderia cenocepacia J2315, an Epidemic Pathogen of Cystic Fibrosis Patients

Genome-Wide Expression Analysis of Burkholderia pseudomallei Infection ina Hamster Model of Acute Melioidosis

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