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Applied and Environmental Microbiology, October 2001, p. 4560-4565, Vol. 67, No. 10
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.10.4560-4565.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Mutations in the Listerial proB Gene Leading to Proline Overproduction: Effects on Salt Tolerance and Murine Infection

Roy D. Sleator, Cormac G. M. Gahan, and Colin Hill*

Department of Microbiology and National Food Biotechnology Centre, University College Cork, Cork, Ireland

Received 5 March 2001/Accepted 16 July 2001

The observed sensitivity of Listeria monocytogenes to the toxic proline analogue L-azetidine-2-carboxylic acid (AZ) suggested that proline synthesis in Listeria may be regulated by feedback inhibition of gamma -glutamyl kinase (GK), the first enzyme of the proline biosynthesis pathway, encoded by the proB gene. Taking advantage of the Epicurian coli mutator strain XL1-Red, we performed random mutagenesis of the recently described proBA operon and generated three independent mutations in the listerial proB homologue, leading to proline overproduction and salt tolerance when expressed in an E. coli (Delta proBA) background. While each of the mutations (located within a conserved 26-amino-acid region of GK) was shown to confer AZ resistance (AZr) on an L. monocytogenes proBA mutant, listerial transformants failed to exhibit the salt-tolerant phenotype observed in E. coli. Since proline accumulation has previously been linked to the virulence potential of a number of pathogenic bacteria, we analyzed the effect of proline overproduction on Listeria pathogenesis. However, our results suggest that as previously described for proline auxotrophy, proline hyperproduction has no apparent impact on the virulence potential of Listeria.

* Corresponding author. Mailing address: Department of Microbiology, University College Cork, Cork, Ireland. Phone: 353-21-4902397. Fax: 353-21-4903101. E-mail: c.hill{at}exchsvrs.ucc.ie.

Applied and Environmental Microbiology, October 2001, p. 4560-4565, Vol. 67, No. 10
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.10.4560-4565.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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  • Kim, S. H., Bakko, M. K., Knowles, D., Borucki, M. K. (2004). Oral Inoculation of A/J Mice for Detection of Invasiveness Differences between Listeria monocytogenes Epidemic and Environmental Strains. Infect. Immun. 72: 4318-4321 [Abstract] [Full Text]  
  • Fujita, T., Maggio, A., Garcia-Rios, M., Stauffacher, C., Bressan, R. A., Csonka, L. N. (2003). Identification of Regions of the Tomato gamma -Glutamyl Kinase That Are Involved in Allosteric Regulation by Proline. J. Biol. Chem. 278: 14203-14210 [Abstract] [Full Text]  
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