Absence of Serotype-Specific Surface Antigen and Altered Teichoic Acid Glycosylation among Epidemic-Associated  Strains of Listeria monocytogenes

Outbreaks of food-borne listeriosis have often involved strains of serotype 4b. Examination of multiple isolates from three different outbreaks revealed that ca. 11 to 29% of each epidemic population consisted of strains which were negative with the serotype-specific monoclonal antibody c74.22, lacked galactose from the teichoic acid of the cell wall, and were resistant to the serotype 4b-specific phage 2671.

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Absence of Serotype-Specific Surface Antigen in Laboratory Variants of Epidemic-Associated Listeria monocytogenes Strains

Applied and Environmental Microbiology ◽

10.1128/aem.00473-07 ◽

2007 ◽

Vol 73 (19) ◽

pp. 6313-6316 ◽

Cited By ~ 5

Author(s):

Ying Cheng ◽

Lili Yue ◽

Driss Elhanafi ◽

S. Kathariou

Keyword(s):

Monoclonal Antibody ◽

Listeria Monocytogenes ◽

Specific Surface ◽

Surface Antigen ◽

Specific Monoclonal Antibody ◽

Genetic Studies ◽

Laboratory Conditions ◽

Serotype 4B ◽

Genetic Constructs

ABSTRACT Variants that lacked reactivity with the serotype 4b-specific monoclonal antibody c74.22 and that lost susceptibility to certain Listeria- or serotype 4b-specific phages were identified in the course of genetic studies with serotype 4b Listeria monocytogenes strains H7550 and F2381L (epidemic clones I and II, respectively). Our findings suggest that such variants can become inadvertently established under laboratory conditions and suggest caution in work involving serotype 4b strains and genetic constructs thereof.

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Teichoic Acid Glycosylation Mediated by gtcA Is Required for Phage Adsorption and Susceptibility of Listeria monocytogenes Serotype 4b

Applied and Environmental Microbiology ◽

10.1128/aem.01773-07 ◽

2008 ◽

Vol 74 (5) ◽

pp. 1653-1655 ◽

Cited By ~ 25

Author(s):

Ying Cheng ◽

Nattawan Promadej ◽

Jae-Won Kim ◽

S. Kathariou

Keyword(s):

Cell Wall ◽

Listeria Monocytogenes ◽

Teichoic Acid ◽

Insertion Mutant ◽

Wall Teichoic Acid ◽

Phage Adsorption ◽

Serotype 4B

ABSTRACT An insertion mutant of gtcA, responsible for serotype-specific glycosylation of the cell wall teichoic acid in serotype 4b strains of Listeria monocytogenes, was also resistant to both Listeria genus- and serotype 4b-specific phages. The sugar substituents on teichoic acid appeared essential for the adsorption of phages A500 (serotype 4b specific) and A511 (Listeria genus specific) to serotype 4b L. monocytogenes.

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A Sheep in Wolf's Clothing: Listeria innocua Strains with Teichoic Acid-Associated Surface Antigens and Genes Characteristic of Listeria monocytogenes Serogroup 4

Journal of Bacteriology ◽

10.1128/jb.182.21.6161-6168.2000 ◽

2000 ◽

Vol 182 (21) ◽

pp. 6161-6168 ◽

Cited By ~ 25

Author(s):

Zheng Lan ◽

Franz Fiedler ◽

Sophia Kathariou

Keyword(s):

Listeria Monocytogenes ◽

Genomic Organization ◽

Teichoic Acid ◽

Surface Antigens ◽

Listeria Innocua ◽

Genetic Studies ◽

Food Borne ◽

Antigenic Properties ◽

Serotype 4B ◽

High Degree

ABSTRACT Listeria monocytogenes serotype 4b has been implicated in numerous food-borne epidemics and in a substantial fraction of sporadic listeriosis. A unique lineage of the nonpathogenic speciesListeria innocua was found to express teichoic acid-associated surface antigens that were otherwise expressed only byL. monocytogenes of serotype 4b and the rare serotypes 4d and 4e. These L. innocua strains were also found to harbor sequences homologous to the gene gtcA, which has been shown to be essential for teichoic acid glycosylation inL. monocytogenes serotype 4b. Transposon mutagenesis and genetic studies revealed that the gtcA gene identified in this lineage of L. innocua was functional in serotype 4b-like glycosylation of the teichoic acids of these organisms. The genomic organization of the gtcA region was conserved between this lineage of L. innocua and L. monocytogenes serotype 4b. Our data are in agreement with the hypothesis that, in this lineage of L. innocua,gtcA was acquired by lateral transfer from L. monocytogenes serogroup 4. The high degree of nucleotide sequence conservation in the gtcA sequences suggests that such transfer was relatively recent. Transfer events of this type may alter the surface antigenic properties of L. innocua and may eventually lead to evolution of novel pathogenic lineages through additional acquisition of genes from virulent listeriae.

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Cell Wall Teichoic Acid Glycosylation inListeria monocytogenes Serotype 4b Requires gtcA, a Novel, Serogroup-Specific Gene

Journal of Bacteriology ◽

10.1128/jb.181.2.418-425.1999 ◽

1999 ◽

Vol 181 (2) ◽

pp. 418-425 ◽

Cited By ~ 63

Author(s):

N. Promadej ◽

F. Fiedler ◽

P. Cossart ◽

S. Dramsi ◽

S. Kathariou

Keyword(s):

Cell Wall ◽

Marked Reduction ◽

Teichoic Acid ◽

Lipoteichoic Acid ◽

Specific Gene ◽

Specific Monoclonal Antibody ◽

Wild Type ◽

Wall Teichoic Acid ◽

Insertional Inactivation ◽

Serotype 4B

ABSTRACT We have identified a novel gene, gtcA, involved in the decoration of cell wall teichoic acid of Listeria monocytogenes serotype 4b with galactose and glucose. Insertional inactivation of gtcA brought about loss of reactivity with the serotype 4b-specific monoclonal antibody c74.22 and was accompanied by a complete lack of galactose and a marked reduction in the amounts of glucose on teichoic acid. Interestingly, the composition of membrane-associated lipoteichoic acid was not affected. Complementation of the mutants with the clonedgtcA in trans restored galactose and glucose on teichoic acid to wild-type levels. The complemented strains also recovered reactivity with c74.22. Within L. monocytogenes, sequences homologous to gtcA were found in all serogroup 4 isolates but not in strains of any other serotypes. In serotype 4b, gtcA appears to be the first member of a bicistronic operon which includes a gene with homology toBacillus subtilis rpmE, encoding ribosomal protein L31. In contrast to gtcA, the latter gene appears conserved among all screened serotypes of L. monocytogenes.

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Construction, Characterization, and Use of Two Listeria monocytogenes Site-Specific Phage Integration Vectors

Journal of Bacteriology ◽

10.1128/jb.184.15.4177-4186.2002 ◽

2002 ◽

Vol 184 (15) ◽

pp. 4177-4186 ◽

Cited By ~ 320

Author(s):

Peter Lauer ◽

Man Yin Nora Chow ◽

Martin J. Loessner ◽

Daniel A. Portnoy ◽

Richard Calendar

Keyword(s):

Listeria Monocytogenes ◽

Lethal Dose ◽

Attachment Site ◽

Donor Cell ◽

Single Copy ◽

Cell Extract ◽

Site Specific ◽

Specific Phage ◽

Serotype 1 ◽

Serotype 4B

ABSTRACT Two site-specific shuttle integration vectors were developed with two different chromosomal bacteriophage integration sites to facilitate strain construction in Listeria monocytogenes. The first vector, pPL1, utilizes the listeriophage U153 integrase and attachment site within the comK gene for chromosomal insertion. pPL1 contains a useful polylinker, can be directly conjugated from Escherichia coli into L. monocytogenes, forms stable, single-copy integrants at a frequency of ∼10−4 per donor cell, and can be used in the L. monocytogenes 1/2 and 4b serogroups. Methods for curing endogenous prophages from the comK attachment site in 10403S-derived strains were developed. pPL1 was used to introduce the hly and actA genes at comK-attBB′ in deletion strains derived from 10403S and SLCC-5764. These strains were tested for second-site complementation in hemolysin assays, plaquing assays, and cell extract motility assays. Unlike plasmid-complemented strains, integrated pPL1-complemented strains were fully virulent in the mouse 50% lethal dose assay. Additionally, the PSA phage attachment site on the L. monocytogenes chromosome was characterized, and pPL1 was modified to integrate at this site. The listeriophage PSA integrates in the 3′ end of an arginine tRNA gene. There are 17 bp of DNA identity between the bacterial and phage attachment sites. The PSA prophage DNA sequence reconstitutes a complete tRNAArg gene. The modified vector, pPL2, was integration proficient at the same frequency as pPL1 in common laboratory serotype 1/2 strains as well as serotype 4b strains.

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Conservation of Genomic Localization and Sequence Content of Sau3AI-Like Restriction-Modification Gene Cassettes among Listeria monocytogenes Epidemic Clone I and Selected Strains of Serotype 1/2a

Applied and Environmental Microbiology ◽

10.1128/aem.00648-10 ◽

2010 ◽

Vol 76 (16) ◽

pp. 5577-5584 ◽

Cited By ~ 11

Author(s):

Suleyman Yildirim ◽

Driss Elhanafi ◽

Wen Lin ◽

Anthony D. Hitchins ◽

Robin M. Siletzky ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Gene Cassette ◽

Conclusive Evidence ◽

Gene Encoding ◽

Epidemic Clone ◽

Food Borne ◽

Serotype 1 ◽

Serotype 4B ◽

Clonal Population Structure ◽

Restriction Modification

ABSTRACTListeria monocytogenesis a food-borne pathogen with a clonal population structure and apparently limited gene flow between strains of different lineages. Strains of epidemic clone I (ECI) have been responsible for numerous outbreaks and invariably have DNA that is resistant to digestion by Sau3AI, suggesting methylation of cytosine at GATC sites. A putative restriction-modification (RM) gene cassette has been identified in the genome of the ECI strain F2365 and all other tested ECI strains but is absent from other strains of the same serotype (4b). Homologous RM cassettes have not been reported amongL. monocytogenesisolates of other serotypes. Furthermore, conclusive evidence for the involvement of this RM cassette in the Sau3AI resistance phenotype of ECI strains has been lacking. In this study, we describe a highly conserved RM cassette in certain strains of serotypes 1/2a and 4a that have Sau3AI-resistant DNA. In these strains the RM cassette was in the same genomic location as in the ECI reference strain F2365. The cassette included a gene encoding a putative recombinase, suggesting insertion via site-specific recombination. Deletion of the RM cassette in the ECI strain F2365 and the serotype 1/2a strain A7 rendered the DNA of both strains susceptible to Sau3AI digestion, providing conclusive evidence that the cassette includes a gene required for methylation of cytosine at GATC sites in both strains. The findings suggest that, in addition to its presence in ECI strains, this RM cassette and the accompanying genomic DNA methylation is also encountered among selected strains of other lineages.

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Comparative Proteomic Analysis of Listeria monocytogenes Strains F2365 and EGD

Applied and Environmental Microbiology ◽

10.1128/aem.01847-08 ◽

2008 ◽

Vol 75 (2) ◽

pp. 366-373 ◽

Cited By ~ 33

Author(s):

Janet R. Donaldson ◽

Bindu Nanduri ◽

Shane C. Burgess ◽

Mark L. Lawrence

Keyword(s):

Listeria Monocytogenes ◽

Protein Identification ◽

Altered Expression ◽

Genetic Lineages ◽

Multidimensional Protein Identification Technology ◽

Food Borne ◽

Flagellar Biosynthesis ◽

Serotype 1 ◽

Serotype 4B ◽

Biological Differences

ABSTRACT Listeria monocytogenes is a gram-positive, food-borne pathogen that causes disease in both humans and animals. There are three major genetic lineages of L. monocytogenes and 13 serovars. To further our understanding of the differences that exist between different genetic lineages/serovars of L. monocytogenes, we analyzed the global protein expression of the serotype 1/2a strain EGD and the serotype 4b strain F2365 during early-stationary-phase growth at 37°C. Using multidimensional protein identification technology with electrospray ionization tandem mass spectrometry, we identified 1,754 proteins from EGD and 1,427 proteins from F2365, of which 1,077 were common to both. Analysis of proteins that had significantly altered expression between strains revealed potential biological differences between these two L. monocytogenes strains. In particular, the strains differed in expression of proteins involved in cell wall physiology and flagellar biosynthesis, as well as DNA repair proteins and stress response proteins.

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