DNA methylation in Yersinia enterocolitica: role of the DNA adenine methyltransferase in mismatch repair and regulation of virulence factors

Fälker, Stefan; Schmidt, Markus; Heusipp, Gerhard

doi:10.1099/mic.0.27946-0

Cited by 29 publications

(50 citation statements)

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“…For routine growth, all strains were grown in LuriaBertani (LB) broth or on agar plates at 26°C for Y. enterocolitica or 37°C for E. coli. Antibiotics were used as described previously (9).…”

Section: Methodsmentioning

confidence: 99%

“…After confirmation of the correct genotype by Southern blotting and PCR, single mutants were used for an additional round of mutagenesis for the construction of pyp double mutants. Finally, pFUSE-hreP was integrated into the chromosome to monitor ␤-galactosidase expression from the hreP-lacZYA chromosomal fusion (9). The correct genotype of each strain was confirmed by Southern blotting and PCR.…”

Section: Methodsmentioning

confidence: 99%

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A Regulatory Network Controls Expression of the In Vivo-Expressed HreP Protease of Yersinia enterocolitica

et al. 2009

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The human enteropathogen Yersinia enterocolitica survives and replicates in the lymphoid tissues of its host. Previous in vivo analyses of gene expression revealed that various chromosomal genes are expressed at this stage of infection, but not in vitro. One of these, termed hreP, encodes a protease that is necessary for full virulence of Y. enterocolitica. Using transposon mutagenesis, we identified three genes, pypA, pypB, and pypC, as positive regulators of hreP transcription. PypA is an inner membrane protein with no significant similarity to any known proteins; PypB is a ToxR-like transmembrane transcriptional regulator; and PypC is a cytoplasmic transcriptional regulator with an OmpR-like winged helix-turn-helix DNA binding motif. We show that all Pyp proteins are able to activate hreP independently of each other and that PypB and PypC interact directly with the hreP promoter region. Furthermore, pypB and pypC are autoregulated and regulate each other. Additional data indicate that transcription of hreP is repressed by the histone-like nucleoid-structuring protein H-NS in a temperature-dependent manner. Our data reveal a new regulatory network that might have implications for the controlled expression of further virulence-associated functions in Yersinia.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

A Regulatory Network Controls Expression of the In Vivo-Expressed HreP Protease of Yersinia enterocolitica

et al. 2009

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“…Unless otherwise indicated, all strains were grown in Luria-Bertani (LB) broth or on agar plates at 26°C for Y. enterocolitica or 37°C for E. coli. Antibiotics were used as described previously (20).…”

Section: Methodsmentioning

confidence: 99%

“…For the construction of Y. enterocolitica promoter-lacZ fusions, ϳ600-bp fragments containing the 5Ј end of the respective gene, including the promoter region, were amplified by PCR using the primer pairs SF-lcrG3/SF-lcrG4, SF-yopN3/SF-yopN4, SF-yscN1/SF-yscN2, SF-orf76-1/ SF-orf76-2, and SF-orf83-1/SF-orf83-2 (see Table 2), with genomic DNA of Y. enterocolitica JB580v as the template. The PCR products were ligated into pKN8 (20), resulting in pKN8-lcrG, pKN8-yopN, pKN8-yscN, pKN8-orf76, and pKN8-orf83. After conjugation of the plasmids to Y. enterocolitica JB580v from E. coli S17-1pir and integration into the virulence plasmid, merodiploid lcrG-lacZYA/ lcrG into the Y. enterocolitica virulence plasmid was confirmed by Southern blot analysis (data not shown).…”

Section: Methodsmentioning

confidence: 99%

“…(Fig. 1), which are involved in the control of Yop release and have been described as "gatekeepers" of Yop secretion (10,15,21,31,51,63 (20). Therefore, translocated as well as bacterium-associated Yop proteins could potentially be detected in the cytosol of host cells after immunoprecipitation with Yop-specific antisera, and this might be a reason for the differences in translocation between Dam OP and Ca 2ϩ -blind strains.…”

Section: Methodsmentioning

confidence: 99%

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Altered Ca²⁺Regulation of Yop Secretion inYersinia enterocoliticaafter DNA Adenine Methyltransferase Overproduction Is Mediated by Clp-Dependent Degradation of LcrG

2006

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In Escherichia coli, the Dam enzyme (DNA adenine methyltransferase) catalyzes the methylation of adenine residues at the N-6 position in GATC sequences. The methylation of DNA influences various basic cellular processes ranging from chromosome replication to mismatch repair (72). During replication, delayed methylation of the newly synthesized daughter strand is required for parental strand-directed mismatch repair (48). Therefore, an imbalance in DNA methylation in dam mutant or Dam-overproducing (Dam OP ) strains results in an increased mutation frequency and sensitivity to base analogues (23,28,45). In addition to having these functions, DNA methylation influences the binding of regulatory proteins to promoter regions, thereby imparting an epigenetic mechanism linked to replication on gene expression. The best-studied example is the expression of pyelonephritis-associated pili (pap) of uropathogenic E. coli, where Dam controls the binding of Lrp in the promoter region of the pap operon (29).Furthermore, Dam has been shown to be essential for virulence in a number of human and animal pathogens, like Salmonella enterica, Yersinia pseudotuberculosis, Yersinia pestis, Haemophilus influenzae, Vibrio cholerae, Pasteurella multocida, and Aeromonas hydrophila (9,19,36,37,71 (3,36,37). Attenuation of these strains was used effectively in the development of vaccine strains of Salmonella as well as Yersinia (17,18,26,36,57,66). The phenotypes of strains with defects in DNA methylation are generally diverse, which is not surprising given the fact that GATC sequences are widespread in the chromosome. Interestingly, effects of Dam on the secretion of virulence factors seem to be common among pathogens. Dam OP interferes with the regulation of type III secretion (T3S) in S. enterica serovar Typhimurium as well as in Y. pseudotuberculosis (22,36,37). Furthermore, dam mutants of S. enterica serovar Typhimurium release large amounts of membrane material containing extracellular proteins into the supernatant (55). In A. hydrophila, the T3S-associated cytotoxicity of a Dam OP strain is decreased, while the cytotoxicity associated with the type II secreted Act enterotoxin is increased (19). The regulatory networks behind these phenomena, however, remain elusive. Although Dam OP strains as well as dam mutant strains might not reflect a physiologically relevant situation, they provide excellent models for studying the influence of DNA methylation on gene expression. Therefore, employing such strains is primarily a tool for studying effects of differential DNA methylation in promoter regions that are of physiological significance (72). This does not imply a direct regulation by Dam but mirrors the effect of altered binding affinities of methylation-sensitive regulatory proteins.The virulence plasmid-encoded T3S system (T3SS) is a hallmark of Yersinia virulence. Upon host cell contact, the tightly regulated Yop/Ysc T3SS is expressed and translocates the Yop effector proteins into the host cell cytosol, where they downregulate the hos...

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Epigenetic Programming by Microbial Pathogens and Impacts on Acute and Chronic Disease

Mahan

Heithoff

Barnes

et al. 2017

Epigenetics of Infectious Diseases

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DNA methylation in Yersinia enterocolitica: role of the DNA adenine methyltransferase in mismatch repair and regulation of virulence factors

Cited by 29 publications

References 43 publications

A Regulatory Network Controls Expression of the In Vivo-Expressed HreP Protease of Yersinia enterocolitica

A Regulatory Network Controls Expression of the In Vivo-Expressed HreP Protease of Yersinia enterocolitica

Altered Ca²⁺Regulation of Yop Secretion inYersinia enterocoliticaafter DNA Adenine Methyltransferase Overproduction Is Mediated by Clp-Dependent Degradation of LcrG

Epigenetic Programming by Microbial Pathogens and Impacts on Acute and Chronic Disease

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DNA methylation in Yersinia enterocolitica: role of the DNA adenine methyltransferase in mismatch repair and regulation of virulence factors

Cited by 29 publications

References 43 publications

A Regulatory Network Controls Expression of the In Vivo-Expressed HreP Protease of Yersinia enterocolitica

A Regulatory Network Controls Expression of the In Vivo-Expressed HreP Protease of Yersinia enterocolitica

Altered Ca2+Regulation of Yop Secretion inYersinia enterocoliticaafter DNA Adenine Methyltransferase Overproduction Is Mediated by Clp-Dependent Degradation of LcrG

Epigenetic Programming by Microbial Pathogens and Impacts on Acute and Chronic Disease

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Altered Ca²⁺Regulation of Yop Secretion inYersinia enterocoliticaafter DNA Adenine Methyltransferase Overproduction Is Mediated by Clp-Dependent Degradation of LcrG