CcpA Regulates Arginine Biosynthesis in Staphylococcus aureus through Repression of Proline Catabolism

Authors

Austin S. Nuxoll, University of Nebraska Medical Center
Steven M. Halouska, University of Nebraska-Lincoln
Marat R. Sadykov, University of Nebraska Medical Center
Mark L. Hanke, University of Nebraska Medical Center
Kenneth W. Bayles, University of Nebraska Medical CenterFollow
Tammy Kielian, University of Nebraska Medical Center
Robert Powers, University of Nebraska-LincolnFollow
Paul D. Fey, University of Nebraska Medical CenterFollow

ORCID IDs

Kenneth W. Bayles

Document Type

Article

Date of this Version

2012

Citation

Nuxoll AS, Halouska SM, Sadykov MR, Hanke ML, Bayles KW, et al. (2012) CcpA Regulates Arginine Biosynthesis in Staphylococcus aureus through Repression of Proline Catabolism. PLoS Pathog 8(11): e1003033. doi:10.1371/journal.ppat.1003033

Comments

Copyright 2012 Nuxoll et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License.

Abstract

Staphylococcus aureus is a leading cause of community-associated and nosocomial infections. Imperative to the success of S. aureus is the ability to adapt and utilize nutrients that are readily available. Genomic sequencing suggests that S. aureus has the genes required for synthesis of all twenty amino acids. However, in vitro experimentation demonstrates that staphylococci have multiple amino acid auxotrophies, including arginine. Although S. aureus possesses the highly conserved anabolic pathway that synthesizes arginine via glutamate, we demonstrate here that inactivation of ccpA facilitates the synthesis of arginine via the urea cycle utilizing proline as a substrate. Mutations within putA, rocD, arcB1, argG and argH abolished the ability of S. aureus JE2 ccpA::tetL to grow in the absence of arginine, whereas an interruption in argJBCF, arcB2, or proC had no effect. Furthermore, nuclear magnetic resonance demonstrated that JE2 ccpA::ermB produced ¹³C₅ labeled arginine when grown with ¹³C₅ proline. Taken together, these data support the conclusion that S. aureus synthesizes arginine from proline during growth on secondary carbon sources. Furthermore, although highly conserved in all sequenced S. aureus genomes, the arginine anabolic pathway (ArgJBCDFGH) is not functional under in vitro growth conditions. Finally, a mutation in argH attenuated virulence in a mouse kidney abscess model in comparison to wild type JE2 demonstrating the importance of arginine biosynthesis in vivo via the urea cycle. However, mutations in argB, argF, and putA did not attenuate virulence suggesting both the glutamate and proline pathways are active and they, or their pathway intermediates, can complement each other in vivo.