Listeria monocytogenes Mutant Defective in Bacteriophage Attachment Is Attenuated in Orally Inoculated Mice and Impaired in Enterocyte Intracellular Growth

A Listeria monocytogenes bacteriophage was used to identify a phage-resistant Tn917 insertion mutant of the mouse-virulent listerial strain F6214-1. The mutant was attenuated when it was inoculated orally into female A/J mice and failed to replicate efficiently in cultured mouse enterocytes. Phage b...

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Bibliographic Details
Published in:Infection and Immunity 2008-09, Vol.76 (9), p.4046-4054
Main Authors: Spears, Patricia A, Suyemoto, M. Mitsu, Palermo, Angela M, Horton, John R, Hamrick, Terri S, Havell, Edward A, Orndorff, Paul E
Format: Article
Language:English
Subjects:
Animals
Bacteria
Bacterial Adhesion
Bacterial Infections
Bacteriology
Bacteriophages - physiology
Biological and medical sciences
DNA Transposable Elements
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
Enterocytes - microbiology
Female
Fundamental and applied biological sciences. Psychology
Genetic Complementation Test
Glycosyltransferases - genetics
Listeria innocua
Listeria monocytogenes
Listeria monocytogenes - pathogenicity
Listeria monocytogenes - virology
Listeria welshimeri
Listeriosis - microbiology
Liver - microbiology
Mice
Mice, Inbred A
Microbiology
Miscellaneous
Molecular Sequence Data
Mutagenesis, Insertional
Receptors, Virus - genetics
Sequence Analysis, DNA
Spleen - microbiology
Survival Analysis
Viral Plaque Assay
Virology
Virulence
Virulence Factors - genetics
Virus Attachment
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Summary:A Listeria monocytogenes bacteriophage was used to identify a phage-resistant Tn917 insertion mutant of the mouse-virulent listerial strain F6214-1. The mutant was attenuated when it was inoculated orally into female A/J mice and failed to replicate efficiently in cultured mouse enterocytes. Phage binding studies indicated that the mutant had a cell surface alteration that precluded phage attachment. All phenotypes associated with the mutation could be complemented in trans by a single open reading frame (ORF) that corresponded to the ORF interrupted by the Tn917 insertion. The complementation effected was, in all cases, at a level indistinguishable from that of the parent. The Tn917 insertion interrupted a gene that is predicted to encode a group 2 glycosyl transferase (provisionally designated glcV). A similar glcV gene is present in Listeria welshimeri and Listeria innocua and in some serotypes of L. monocytogenes. We speculate that the loss of the glcV product results in a defective phage receptor and that this alteration coincidentally influences a feature of the normal host-pathogen interaction required for virulence. Interestingly, the glcV lesion, while preventing phage attachment, did not alter the mutant's ability to bind to cultured mouse enterocyte monolayers. Rather, the mutation appeared to alter a subsequent step in intracellular replication measured by a reduction in plaque-forming efficiency and plaque size. In vivo, the mutant was undetectable in the liver and spleen 48 h after oral inoculation. The mutation is significant in part because it is one of the few that produce attenuation when the mutant is delivered orally.
ISSN:0019-9567
1098-5522
DOI:10.1128/IAI.00283-08