Pyruvate Carboxylase Plays a Crucial Role in Carbon Metabolism of Extra- and Intracellularly Replicating Listeria Monocytogenes

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 2010 Jan 26

PMID 20097852

Citations 38

Authors

Jennifer Schar

Regina Stoll

Kristina Schauer

Daniela I M Loeffler

Eva Eylert

Biju Joseph

Wolfgang Eisenreich

Thilo M Fuchs

Werner Goebel

Affiliations

Soon will be listed here.

Abstract

The human pathogen L. monocytogenes is a facultatively intracellular bacterium that survives and replicates in the cytosol of many mammalian cells. The listerial metabolism, especially under intracellular conditions, is still poorly understood. Recent studies analyzed the carbon metabolism of L. monocytogenes by the (13)C isotopologue perturbation method in a defined minimal medium containing [U-(13)C(6)]glucose. It was shown that these bacteria produce oxaloacetate mainly by carboxylation of pyruvate due to an incomplete tricarboxylic acid cycle. Here, we report that a pycA insertion mutant defective in pyruvate carboxylase (PYC) still grows, albeit at a reduced rate, in brain heart infusion (BHI) medium but is unable to multiply in a defined minimal medium with glucose or glycerol as a carbon source. Aspartate and glutamate of the pycA mutant, in contrast to the wild-type strain, remain unlabeled when [U-(13)C(6)]glucose is added to BHI, indicating that the PYC-catalyzed carboxylation of pyruvate is the predominant reaction leading to oxaloacetate in L. monocytogenes. The pycA mutant is also unable to replicate in mammalian cells and exhibits high virulence attenuation in the mouse sepsis model.

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