Catabolic Ornithine Carbamoyltransferase Activity Facilitates Growth of Staphylococcus aureus in Defined Medium Lacking Glucose and Arginine

Authors

Itidal Reslane, University of Nebraska Medical CenterFollow
Cortney R. Halsey, University of Nebraska Medical Center
Amanda Stastny, University of Nebraska Medical CenterFollow
Barbara J. Cabrera, University of Nebraska Medical CenterFollow
Jong-Sam Ahn, University of Nebraska Medical CenterFollow
Dhananjay Shinde, University of Nebraska Medical CenterFollow
Madeline R. Galac, Walter Reed Army Institute of Research
Margaret F. Sladek, University of Nebraska Medical CenterFollow
Fareha Razvi, University of Nebraska Medical CenterFollow
McKenzie K. Lehman, University of Nebraska Medical CenterFollow
Kenneth W. Bayles, University of Nebraska Medical CenterFollow
Vinai Chittezham Thomas, University of Nebraska Medical CenterFollow
Luke D. Handke, University of Nebraska Medical CenterFollow
Paul D. Fey, University of Nebraska Medical CenterFollow

Document Type

Article

Journal Title

mBio

Publication Date

2022

Volume

13

Abstract

Previous studies have found that arginine biosynthesis in Staphylococcus aureus is repressed via carbon catabolite repression (CcpA), and proline is used as a precursor. Unexpectedly, however, robust growth of S. aureus is not observed in complete defined medium lacking both glucose and arginine (CDM-R). Mutants able to grow on agar-containing defined medium lacking arginine (CDM-R) were selected and found to contain mutations within ahrC, encoding the canonical arginine biosynthesis pathway repressor (AhrC), or single nucleotide polymorphisms (SNPs) upstream of the native arginine deiminase (ADI) operon arcA1B1D1C1. Reverse transcription-PCR (RT-PCR) studies found that mutations within ccpA or ahrC or SNPs identified upstream of arcA1B1D1C1 increased the transcription of both arcB1 and argGH, encoding ornithine carbamoyltransferase and argininosuccinate synthase/lyase, respectively, facilitating arginine biosynthesis. Furthermore, mutations within the AhrC homologue argR2 facilitated robust growth within CDM-R. Complementation with arcB1 or arcA1B1D1C1, but not argGH, rescued growth in CDM-R. Finally, supplementation of CDM-R with ornithine stimulated growth, as did mutations in genes (proC and rocA) that presumably increased the pyrroline-5-carboxylate and ornithine pools. Collectively, these data suggest that the transcriptional regulation of ornithine carbamoyltransferase and, in addition, the availability of intracellular ornithine pools regulate arginine biosynthesis in S. aureus in the absence of glucose. Surprisingly, ~50% of clinical S. aureus isolates were able to grow in CDM-R. These data suggest that S. aureus is selected to repress arginine biosynthesis in environments with or without glucose; however, mutants may be readily selected that facilitate arginine biosynthesis and growth in specific environments lacking arginine. IMPORTANCE Staphylococcus aureus can cause infection in virtually any niche of the human host, suggesting that it has significant metabolic versatility. Indeed, bioinformatic analysis suggests that it has the biosynthetic capability to synthesize all 20 amino acids. Paradoxically, however, it is conditionally auxotrophic for several amino acids, including arginine. Studies in our laboratory are designed to assess the biological function of amino acid auxotrophy in this significant pathogen. This study reveals that the metabolic block repressing arginine biosynthesis in media lacking glucose is the transcriptional repression of ornithine carbamoyltransferase encoded by arcB1 within the native arginine deiminase operon in addition to limited intracellular pools of ornithine. Surprisingly, approximately 50% of S. aureus clinical isolates can grow in media lacking arginine, suggesting that mutations are selected in S. aureus that allow growth in particular niches of the human host.

MeSH Headings

Amino Acids, Arginine, Bacterial Proteins, Glucose, Ornithine, Ornithine Carbamoyltransferase, Staphylococcus aureus

DOI Link

10.1128/mbio.00395-22

ISSN

2150-7511

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Reslane, Itidal; Halsey, Cortney R.; Stastny, Amanda; Cabrera, Barbara J.; Ahn, Jong-Sam; Shinde, Dhananjay; Galac, Madeline R.; Sladek, Margaret F.; Razvi, Fareha; Lehman, McKenzie K.; Bayles, Kenneth W.; Chittezham Thomas, Vinai; Handke, Luke D.; and Fey, Paul D., "Catabolic Ornithine Carbamoyltransferase Activity Facilitates Growth of Staphylococcus aureus in Defined Medium Lacking Glucose and Arginine" (2022). Journal Articles: Pathology and Microbiology. 88.
https://digitalcommons.unmc.edu/com_pathmicro_articles/88