Cloning, expression, and purification of the virulence-associated protein D from Xylella fastidiosa

Catani, Cleide Ferreira; Azzoni, Adriano Rodrigues; Paula, Danielli Parrilha de; Tada, Susely F.S.; Rosselli, Luciana Kauer; Souza, Anete Pereira de; Yano, Tomomasa

doi:10.1016/j.pep.2004.07.002

Cited by 9 publications

(9 citation statements)

References 27 publications

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“…Interestingly the genetic load region also contains the vapD gene that encodes the virulence associated protein D. VapD was shown to function as a toxin and is generally counteracted by an antitoxin protein called VapX. VapD/VapX systems have been reported in pathogens like Rhodococcus equi [39], Dichelobacter nodosus [40], Xylella fastidiosa [41], and Haemophilus influenzae [42]. It has been shown that expression of VapD enhances the invasion and survival within both human epithelial and endothelial cells [42].…”

Section: Resultsmentioning

confidence: 99%

Conjugative Plasmids of Neisseria gonorrhoeae

Pachulec

Does

2010

PLoS ONE

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Many clinical isolates of the human pathogen Neisseria gonorrhoeae contain conjugative plasmids. The host range of these plasmids is limited to Neisseria species, but presence of a tetracycline (tetM) determinant inserted in several of these plasmids is an important cause of the rapid spread of tetracycline resistance. Previously plasmids with different backbones (Dutch and American type backbones) and with and without different tetM determinants (Dutch and American type tetM determinants) have been identified. Within the isolates tested, all plasmids with American or Dutch type tetM determinants contained a Dutch type plasmid backbone. This demonstrated that tetM determinants should not be used to differentiate between conjugal plasmid backbones. The nucleotide sequences of conjugative plasmids with Dutch type plasmid backbones either not containing the tetM determinant (pEP5233) or containing Dutch (pEP5289) or American (pEP5050) type tetM determinants were determined. Analysis of the backbone sequences showed that they belong to a novel IncP1 subfamily divergent from the IncP1α, β, γ, δ and ε subfamilies. The tetM determinants were inserted in a genetic load region found in all these plasmids. Insertion was accompanied by the insertion of a gene with an unknown function, and rearrangement of a toxin/antitoxin gene cluster. The genetic load region contains two toxin/antitoxins of the Zeta/Epsilon toxin/antitoxin family previously only found in Gram positive organisms and the virulence associated protein D of the VapD/VapX toxin/antitoxin family. Remarkably, presence of VapX of pJD1, a small cryptic neisserial plasmid, in the acceptor strain strongly increased the conjugation efficiency, suggesting that it functions as an antitoxin for the conjugative plasmid. The presence of the toxin and antitoxin on different plasmids might explain why the host range of this IncP1 plasmid is limited to Neisseria species. The isolated plasmids conjugated efficiently between N. gonorrhoeae strains, but did not enhance transfer of a genetic marker.

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Section: Resultsmentioning

confidence: 99%

Conjugative Plasmids of Neisseria gonorrhoeae

Pachulec

Does

2010

PLoS ONE

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“…However, some predicted virulence factors remain unstudied. One predicted virulence factor of interest was found in the plasmid pXF51; in this plasmid, the ORF XFb0051 encodes the 17.6 kDa virulence-associated protein D (Xf-VapD), which contains a predominance of α-helices in its secondary structure [ 19 ]. Xf-VapD is predicted to belong to the CRISPR/Cas2 family (Pfam: PF09827).…”

Section: Introductionmentioning

confidence: 99%

VapD in Xylella fastidiosa Is a Thermostable Protein with Ribonuclease Activity

et al. 2015

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Xylella fastidiosa strain 9a5c is a gram-negative phytopathogen that is the causal agent of citrus variegated chlorosis (CVC), a disease that is responsible for economic losses in Brazilian agriculture. The most well-known mechanism of pathogenicity for this bacterial pathogen is xylem vessel occlusion, which results from bacterial movement and the formation of biofilms. The molecular mechanisms underlying the virulence caused by biofilm formation are unknown. Here, we provide evidence showing that virulence-associated protein D in X. fastidiosa (Xf-VapD) is a thermostable protein with ribonuclease activity. Moreover, protein expression analyses in two X. fastidiosa strains, including virulent (Xf9a5c) and nonpathogenic (XfJ1a12) strains, showed that Xf-VapD was expressed during all phases of development in both strains and that increased expression was observed in Xf9a5c during biofilm growth. This study is an important step toward characterizing and improving our understanding of the biological significance of Xf-VapD and its potential functions in the CVC pathosystem.

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“…The X. fastidiosa CVC 9a5c strain was found to contain a VapD gene on pXF51 (22). The VapD protein from CVC 9a5c is similar to a VapD found in Riemerella anatipestifer and Actinobacillus actinomycetemcomitans (5). The function of this protein is unknown, although microarray analysis with the CVC 9a5c strain indicates that it may be turned on during heat stress (27).…”

Section: Downloaded Frommentioning

confidence: 99%

Genetic Analysis of a NovelXylella fastidiosaSubspecies Found in the Southwestern United States

Randall

Goldberg

Kemp

et al. 2009

Appl Environ Microbiol

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Xylella fastidiosa, the causal agent of several scorch diseases, is associated with leaf scorch symptoms in Chitalpa tashkentensis, a common ornamental landscape plant used throughout the southwestern United States. For a number of years, many chitalpa trees in southern New Mexico and Arizona exhibited leaf scorch symptoms, and the results from a regional survey show that chitalpa trees from New Mexico, Arizona, and California are frequently infected with X. fastidiosa. Phylogenetic analysis of multiple loci was used to compare the X. fastidiosa infecting chitalpa strains from New Mexico, Arizona, and trees imported into New Mexico nurseries with previously reported X. fastidiosa strains. Loci analyzed included the 16S ribosome, 16S-23S ribosomal intergenic spacer region, gyrase-B, simple sequence repeat sequences, X. fastidiosa-specific sequences, and the virulence-associated protein (VapD). This analysis indicates that the X. fastidiosa isolates associated with infected chitalpa trees in the Southwest are a highly related group that is distinct from the four previously defined taxons X. fastidiosa subsp. fastidiosa (piercei), X. fastidiosa subsp. multiplex, X. fastidiosa subsp. sandyi, and X. fastidiosa subsp. pauca. Therefore, the classification proposed for this new subspecies is X. fastidiosa subsp. tashke.

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Cloning, expression, and purification of the virulence-associated protein D from Xylella fastidiosa

Cited by 9 publications

References 27 publications

Conjugative Plasmids of Neisseria gonorrhoeae

Conjugative Plasmids of Neisseria gonorrhoeae

VapD in Xylella fastidiosa Is a Thermostable Protein with Ribonuclease Activity

Genetic Analysis of a NovelXylella fastidiosaSubspecies Found in the Southwestern United States

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