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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  13 C 5  labeled arginine when grown with  13 C 5  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 .

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