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Infection and Immunity, June 2004, p. 3237-3244, Vol. 72, No. 6
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.6.3237-3244.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Role of Flagellin and the Two-Component CheA/CheY System of Listeria monocytogenes in Host Cell Invasion and Virulence

Lone Dons,1* Emma Eriksson,2 Yuxuan Jin,3 Martin E. Rottenberg,2 Krister Kristensson,3 Charlotte N. Larsen,1,{dagger} José Bresciani,4 and John E. Olsen1

Departments of Veterinary Pathobiology,1 Ecology, The Royal Veterinary and Agricultural University, Frederiksberg C, Denmark,4 Microbiology and Tumor Biology Center,2 Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden3

Received 7 July 2003/ Returned for modification 11 August 2003/ Accepted 26 February 2004

The flagellum protein flagellin of Listeria monocytogenes is encoded by the flaA gene. Immediately downstream of flaA, two genes, cheY and cheA, encoding products with homology to chemotaxis proteins of other bacteria, are located. In this study we constructed deletion mutants with mutations in flaA. cheY, and cheA to elucidate their role in the biology of infection with L. monocytogenes. The {Delta}cheY, {Delta}cheA, and double-mutant {Delta}cheYA mutants, but not {Delta}flaA mutant, were motile in liquid media. However, the {Delta}cheA mutant had impaired swarming and the {Delta}cheY and {Delta}cheYA mutants were unable to swarm on soft agar plates, suggesting that cheY and cheA genes encode proteins involved in chemotaxis. The {Delta}flaA, {Delta}cheY, {Delta}cheA, and {Delta}cheYA mutants (grown at 24°C) showed reduced association with and invasion of Caco-2 cells compared to the wild-type strain. However, spleens from intragastrically infected BALB/c and C57BL/6 mice showed larger and similar numbers of the {Delta}flaA and {Delta}cheYA mutants, respectively, compared to the wild-type controls. Such a discrepancy could be explained by the fact that tumor necrosis factor receptor p55 deficient mice showed dramatically exacerbated susceptibility to the wild-type but unchanged or only slightly increased levels of the {Delta}flaA or {Delta}cheYA mutant. In summary, we show that listerial flaA. cheY, and cheA gene products facilitate the initial contact with epithelial cells and contribute to effective invasion but that flaA could also be involved in the triggering of immune responses.

* Corresponding author. Mailing address: Department of Veterinary Pathobiology, The Royal Veterinary and Agricultural University, Stigbøjlen 4, DK-1870 Frederiksberg C, Denmark. Phone: 45 35 28 37 42. Fax: 45 35 28 27 55. E-mail: Lone.Dons{at}vetmi.kvl.dk.

Editor: V. J. DiRita

{dagger} Present address: Chr. Hansen A/S, 2970 Hørsholm, Denmark.

Infection and Immunity, June 2004, p. 3237-3244, Vol. 72, No. 6
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.6.3237-3244.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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