Insertional inactivation of the Listeria monocytogenes cheYA operon abolishes response to oxygen gradients and reduces the number of flagella

• Published in: Canadian journal of microbiology Vol. 45; no. 8; pp. 646 - 652
• Main Authors: FLANARY, P. L, ALLEN, R. D, DONS, L, KATHARIOU, S
• Format: Journal Article
• Language: English
• Published: Ottawa, ON National Research Council of Canada 01.08.1999; Canadian Science Publishing NRC Research Press
• Subjects: Animals; Bacteria; Bacterial Proteins; Bacteriology; Base Sequence; Biological and medical sciences; Cell Line; Chemotaxis; DNA Transposable Elements; Flagella - physiology; Flagellin - biosynthesis; Flagellin - genetics; Fundamental and applied biological sciences. Psychology; Genes; Genetic Complementation Test; Genetics; Inactivation; Listeria monocytogenes; Listeria monocytogenes - genetics; Listeria monocytogenes - pathogenicity; Listeria monocytogenes - physiology; Membrane Proteins - genetics; Membrane Proteins - metabolism; Methyl-Accepting Chemotaxis Proteins; Mice; Microbiology; Molecular Sequence Data; Mutagenesis, Insertional; Operon - genetics; Oxygen - pharmacology; Pathology; Polymerase Chain Reaction; Sequence Analysis, DNA; Oxygen; Motility; Operon; Insertion mutation; Flagellum; Listeria monocytogenes; Bacteria; Chemotaxis
• ISSN: 0008-4166; 1480-3275
• DOI: 10.1139/cjm-45-8-646

The nucleotide sequence of a region downstream of the Listeria monocytogenes flagellin gene, flaA, revealed two putative chemotaxis genes, cheY and cheA. These genes have been shown to be transcribed as a bicistronic unit. In this study Tn916 delta E mutagenesis was used to generate two mutants, PF10 and PF16, which contain transposon inserts in the promoter region of this operon. These mutants were motile in liquid, but had reduced flagellin expression and were unable to burrow or swarm on soft agar plates. Complementation of the single transposon-copy mutant PF16 with cloned cheY and cheA in trans partially restored microaerotaxis and swarming on soft agar. The complemented strain did not exhibit any increase in flagellin production. Both PF10 and PF16 appear deficient in their ability to attach to the mouse fibroblast cell line 3T3.