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Volume 153, Issue 5

Phenotypic characterization of a virulence-associated protein, VagH, of reveals a tight link between VagH and the type III secretion system Free

Abstract

Recently, a number of attenuated mutants of have been identified using a bioinformatics approach. One of the target genes identified in that study was , which the authors now characterized further. VagH shows homology to HemK of , possessing methyltransferase activity similar to that of HemK, and targeting release factors 1 and 2. Microarray studies comparing the wild-type and the mutant revealed that the mRNA levels of only a few genes were altered in the mutant. By proteome analysis, expression of the virulence determinant YopD was found to be increased, indicating a possible connection between VagH and the virulence plasmid-encoded type III secretion system (T3SS). Further analysis showed that Yop expression and secretion were repressed in a mutant. This phenotype could be suppressed by -complementation with the wild-type gene or by deletion of the negative regulator . Also, in a similar manner to a T3SS-negative mutant, the avirulent mutant was rapidly cleared from Peyer's patches and could not reach the spleen after oral infection of mice. In a manner analogous to that of T3SS mutants, the mutant could not block phagocytosis by macrophages. However, a mutant showed no defects in the T3SS-independent ability to proliferate intracellularly and replicated to levels similar to those of the wild-type in macrophages. In conclusion, the mutant exhibits a virulence phenotype similar to that of a T3SS-negative mutant, indicating a tight link between VagH and type III secretion in .

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Keyword(s): Cy2, 3-(4-carboxymethyl)phenylmethyl-3′-ethyloxacarbocyanine halide N-hydroxysuccinimidyl ester , Cy3, 1-(5-carboxypentyl)-1′-propylindocarbocyanine halide N-hydrosuccinimidyl ester , Cy5, 1-(5-carboxypentyl)-1′-methylindodicarbocyanine halide N-hydroxysuccinimidyl ester , MTase, N5-methyltransferase , RF, release factor , SAM, S-adenosyl-l-methionine and T3SS, type III secretion system
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2007-05-01
2024-04-20
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Phenotypic characterization of a virulence-associated protein, VagH, of Yersinia pseudotuberculosis reveals a tight link between VagH and the type III secretion system
Microbiology 153, 1464 (2007); https://doi.org/10.1099/mic.0.2006/000323-0
/content/journal/micro/10.1099/mic.0.2006/000323-0
/content/journal/micro/10.1099/mic.0.2006/000323-0
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