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Applied and Environmental Microbiology, March 2004, p. 1669-1679, Vol. 70, No. 3
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.3.1669-1679.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

pbp2229-Mediated Nisin Resistance Mechanism in Listeria monocytogenes Confers Cross-Protection to Class IIa Bacteriocins and Affects Virulence Gene Expression

Anne Gravesen,1* Birgitte Kallipolitis,2 Kim Holmstrøm,3 Poul Erik Høiby,3 Manilduth Ramnath,1,4,{dagger} and Susanne Knøchel1

Department of Dairy and Food Science, Centre for Advanced Food Studies, LMC, The Royal Veterinary and Agricultural University, DK-1958 Frederiksberg C,1 Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M,2 Department of Molecular Characterization, Bioneer A/S, DK-2970 Hørsholm, Denmark,3 Department of Biochemistry, University of Stellenbosch, 7602 Matieland, South Africa4

Received 8 August 2003/ Accepted 21 November 2003

It was previously shown that enhanced nisin resistance in some mutants was associated with increased expression of three genes, pbp2229, hpk1021, and lmo2487, encoding a penicillin-binding protein, a histidine kinase, and a protein of unknown function, respectively. In the present work, we determined the direct role of the three genes in nisin resistance. Interruption of pbp2229 and hpk1021 eliminated the nisin resistance phenotype. Interruption of hpk1021 additionally abolished the increase in pbp2229 expression. The results indicate that this nisin resistance mechanism is caused directly by the increase in pbp2229 expression, which in turn is brought about by the increase in hpk1021 expression. We also found a degree of cross-protection between nisin and class IIa bacteriocins and investigated possible mechanisms. The expression of virulence genes in one nisin-resistant mutant and two class IIa bacteriocin-resistant mutants of the same wild-type strain was analyzed, and each mutant consistently showed either an increase or a decrease in the expression of virulence genes (prfA-regulated as well as prfA-independent genes). Although the changes mostly were moderate, the consistency indicates that a mutant-specific change in virulence may occur concomitantly with bacteriocin resistance development.

* Corresponding author. Mailing address: Department of Dairy and Food Science, Centre for Advanced Food Studies, LMC, The Royal Veterinary and Agricultural University, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark. Phone: 45 3528 3272. Fax: 45 3528 3231. E-mail: alg{at}kvl.dk.

{dagger} Present address: BIOCYDEX, IBMIG, University of Poitiers, 86022 Poitiers Cedex, France.

Applied and Environmental Microbiology, March 2004, p. 1669-1679, Vol. 70, No. 3
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.3.1669-1679.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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