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Applied and Environmental Microbiology, December 2009, p. 7310-7318, Vol. 75, No. 23
0099-2240/09/$08.00+0     doi:10.1128/AEM.01366-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Long-Term Survival of Campylobacter jejuni at Low Temperatures Is Dependent on Polynucleotide Phosphorylase Activity{triangledown}

Nabila Haddad,1 Christopher M. Burns,3 Jean Michel Bolla,4 Hervé Prévost,1 Michel Fédérighi,1 Djamel Drider,2 and Jean Michel Cappelier1*

UMR INRA 1014 Sécurité des Aliments et Microbiologie, Ecole Nationale Vétérinaire, Nantes, France,1 UMR INRA 1014 Sécurité des Aliments et Microbiologie, ENITIAA, Nantes, France,2 College of Biomedical Science, Florida Atlantic University, Boca Raton, Florida,3 Transporteurs Membranaires, Chimioresistance et Drug-Design, Université de la Méditerranée UMR MD 1 UFR Médecine/UFR Pharmacie, Marseille, France4

Received 11 June 2009/ Accepted 25 September 2009

Campylobacter jejuni is a leading cause of bacterial gastroenteritis worldwide. Infection generally occurs after ingestion of contaminated poultry products, usually conserved at low temperatures. The mechanisms promoting survival of C. jejuni in the cold remain poorly understood despite several investigations. The present study provides insight into the survival mechanism by establishing the involvement of polynucleotide phosphorylase (PNPase), a 3'-5' exoribonuclease with multiple biological functions in cold survival. The role of PNPase was demonstrated genetically using strains with altered pnp genes (which encode PNPase) created in C. jejuni F38011 and C. jejuni 81-76 backgrounds. Survival assays carried out at low temperatures (4 and 10°C) revealed a difference of 3 log CFU/ml between the wild-type and the pnp deletion ({Delta}pnp) strains. This did not result from a general requirement for PNPase because survival rates of the strains were similar at higher growth temperatures (37 or 42°C). trans-Complementation with plasmid pNH04 carrying the pnp gene under the control of its natural promoter restored the cold survival phenotype to the pnp deletion strains (at 4 and 10°C) but not to the same level as the wild type. In this study we demonstrate the role of PNPase in low-temperature survival of C. jejuni and therefore attribute a novel biological function to PNPase directly related to human health.

* Corresponding author. Mailing address: UMR INRA 1014 Sécurité des aliments et Microbiologie, Ecole Nationale Vétérinaire, Route de Gachet, 44307 Nantes, France. Phone: 33 240687682. Fax: 33 240687762. E-mail: cappelier{at}vet-nantes.fr

{triangledown} Published ahead of print on 2 October 2009.

Applied and Environmental Microbiology, December 2009, p. 7310-7318, Vol. 75, No. 23
0099-2240/09/$08.00+0     doi:10.1128/AEM.01366-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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