CcpA mediates proline auxotrophy and is required for Staphylococcus aureus pathogenesis

Chunling Li, Fei Sun, Hoonsik Cho, Vamshi Yelavarthi, Changmo Sohn, Chuan He, Olaf Schneewind, Taeok Bae

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Human clinical isolates of Staphylococcus aureus, for example, strains Newman and N315, cannot grow in the absence of proline, albeit their sequenced genomes harbor genes for two redundant proline synthesis pathways. We show here that under selective pressure, S. aureus Newman generates proline-prototrophic variants at a frequency of 3 × 10-6, introducing frameshift and missense mutations in ccpA or IS1811 insertions in ptsH, two regulatory genes that carry out carbon catabolite repression (CCR) in staphylococci and other Gram-positive bacteria. S. aureus Newman variants with mutations in rocF (arginase), rocD (ornithine aminotransferase), and proC (Δ1- pyrroline 5-carboxylate [P5C] reductase) are unable to generate proline-prototrophic variants, whereas a variant with a mutation in ocd (ornithine cyclodeaminase) is unaffected. Transposon insertion in ccpA also restored proline prototrophy. CcpA was shown to repress transcription of rocF and rocD, encoding the first two enzymes, but not of proC, encoding the third and final enzyme in the P5C reductase pathway. CcpA bound to the upstream regions of rocF and rocD but not to that of proC. CcpA's binding to the upstream regions was greatly enhanced by phosphorylated HPr. The CCR-mediated proline auxotrophy was lifted when nonpreferred carbohydrates were used as the sole carbon source. The ccpA mutant displayed reduced staphylococcal load and replication in a murine model of staphylococcal abscess formation, indicating that carbon catabolite repression presents an important pathogenesis strategy of S. aureus infections.

Original languageEnglish
Pages (from-to)3883-3892
Number of pages10
JournalJournal of Bacteriology
Volume192
Issue number15
DOIs
StatePublished - Aug 2010

Fingerprint

Proline
Staphylococcus aureus
Catabolite Repression
Pyrroline Carboxylate Reductases
ornithine cyclodeaminase
Ornithine-Oxo-Acid Transaminase
Arginase
Frameshift Mutation
Mutation
Gram-Positive Bacteria
Missense Mutation
Enzymes
Regulator Genes
Staphylococcus
Abscess
Carbon
Carbohydrates
Genome
Infection
Genes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

CcpA mediates proline auxotrophy and is required for Staphylococcus aureus pathogenesis. / Li, Chunling; Sun, Fei; Cho, Hoonsik; Yelavarthi, Vamshi; Sohn, Changmo; He, Chuan; Schneewind, Olaf; Bae, Taeok.

In: Journal of Bacteriology, Vol. 192, No. 15, 08.2010, p. 3883-3892.

Research output: Contribution to journalArticle

Li, C, Sun, F, Cho, H, Yelavarthi, V, Sohn, C, He, C, Schneewind, O & Bae, T 2010, 'CcpA mediates proline auxotrophy and is required for Staphylococcus aureus pathogenesis', Journal of Bacteriology, vol. 192, no. 15, pp. 3883-3892. https://doi.org/10.1128/JB.00237-10
Li, Chunling ; Sun, Fei ; Cho, Hoonsik ; Yelavarthi, Vamshi ; Sohn, Changmo ; He, Chuan ; Schneewind, Olaf ; Bae, Taeok. / CcpA mediates proline auxotrophy and is required for Staphylococcus aureus pathogenesis. In: Journal of Bacteriology. 2010 ; Vol. 192, No. 15. pp. 3883-3892.
@article{fe44368d65044f1caee2e1e628da65ed,
title = "CcpA mediates proline auxotrophy and is required for Staphylococcus aureus pathogenesis",
abstract = "Human clinical isolates of Staphylococcus aureus, for example, strains Newman and N315, cannot grow in the absence of proline, albeit their sequenced genomes harbor genes for two redundant proline synthesis pathways. We show here that under selective pressure, S. aureus Newman generates proline-prototrophic variants at a frequency of 3 × 10-6, introducing frameshift and missense mutations in ccpA or IS1811 insertions in ptsH, two regulatory genes that carry out carbon catabolite repression (CCR) in staphylococci and other Gram-positive bacteria. S. aureus Newman variants with mutations in rocF (arginase), rocD (ornithine aminotransferase), and proC (Δ1- pyrroline 5-carboxylate [P5C] reductase) are unable to generate proline-prototrophic variants, whereas a variant with a mutation in ocd (ornithine cyclodeaminase) is unaffected. Transposon insertion in ccpA also restored proline prototrophy. CcpA was shown to repress transcription of rocF and rocD, encoding the first two enzymes, but not of proC, encoding the third and final enzyme in the P5C reductase pathway. CcpA bound to the upstream regions of rocF and rocD but not to that of proC. CcpA's binding to the upstream regions was greatly enhanced by phosphorylated HPr. The CCR-mediated proline auxotrophy was lifted when nonpreferred carbohydrates were used as the sole carbon source. The ccpA mutant displayed reduced staphylococcal load and replication in a murine model of staphylococcal abscess formation, indicating that carbon catabolite repression presents an important pathogenesis strategy of S. aureus infections.",
author = "Chunling Li and Fei Sun and Hoonsik Cho and Vamshi Yelavarthi and Changmo Sohn and Chuan He and Olaf Schneewind and Taeok Bae",
year = "2010",
month = "8",
doi = "10.1128/JB.00237-10",
language = "English",
volume = "192",
pages = "3883--3892",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "15",

}

TY - JOUR

T1 - CcpA mediates proline auxotrophy and is required for Staphylococcus aureus pathogenesis

AU - Li, Chunling

AU - Sun, Fei

AU - Cho, Hoonsik

AU - Yelavarthi, Vamshi

AU - Sohn, Changmo

AU - He, Chuan

AU - Schneewind, Olaf

AU - Bae, Taeok

PY - 2010/8

Y1 - 2010/8

N2 - Human clinical isolates of Staphylococcus aureus, for example, strains Newman and N315, cannot grow in the absence of proline, albeit their sequenced genomes harbor genes for two redundant proline synthesis pathways. We show here that under selective pressure, S. aureus Newman generates proline-prototrophic variants at a frequency of 3 × 10-6, introducing frameshift and missense mutations in ccpA or IS1811 insertions in ptsH, two regulatory genes that carry out carbon catabolite repression (CCR) in staphylococci and other Gram-positive bacteria. S. aureus Newman variants with mutations in rocF (arginase), rocD (ornithine aminotransferase), and proC (Δ1- pyrroline 5-carboxylate [P5C] reductase) are unable to generate proline-prototrophic variants, whereas a variant with a mutation in ocd (ornithine cyclodeaminase) is unaffected. Transposon insertion in ccpA also restored proline prototrophy. CcpA was shown to repress transcription of rocF and rocD, encoding the first two enzymes, but not of proC, encoding the third and final enzyme in the P5C reductase pathway. CcpA bound to the upstream regions of rocF and rocD but not to that of proC. CcpA's binding to the upstream regions was greatly enhanced by phosphorylated HPr. The CCR-mediated proline auxotrophy was lifted when nonpreferred carbohydrates were used as the sole carbon source. The ccpA mutant displayed reduced staphylococcal load and replication in a murine model of staphylococcal abscess formation, indicating that carbon catabolite repression presents an important pathogenesis strategy of S. aureus infections.

AB - Human clinical isolates of Staphylococcus aureus, for example, strains Newman and N315, cannot grow in the absence of proline, albeit their sequenced genomes harbor genes for two redundant proline synthesis pathways. We show here that under selective pressure, S. aureus Newman generates proline-prototrophic variants at a frequency of 3 × 10-6, introducing frameshift and missense mutations in ccpA or IS1811 insertions in ptsH, two regulatory genes that carry out carbon catabolite repression (CCR) in staphylococci and other Gram-positive bacteria. S. aureus Newman variants with mutations in rocF (arginase), rocD (ornithine aminotransferase), and proC (Δ1- pyrroline 5-carboxylate [P5C] reductase) are unable to generate proline-prototrophic variants, whereas a variant with a mutation in ocd (ornithine cyclodeaminase) is unaffected. Transposon insertion in ccpA also restored proline prototrophy. CcpA was shown to repress transcription of rocF and rocD, encoding the first two enzymes, but not of proC, encoding the third and final enzyme in the P5C reductase pathway. CcpA bound to the upstream regions of rocF and rocD but not to that of proC. CcpA's binding to the upstream regions was greatly enhanced by phosphorylated HPr. The CCR-mediated proline auxotrophy was lifted when nonpreferred carbohydrates were used as the sole carbon source. The ccpA mutant displayed reduced staphylococcal load and replication in a murine model of staphylococcal abscess formation, indicating that carbon catabolite repression presents an important pathogenesis strategy of S. aureus infections.

UR - http://www.scopus.com/inward/record.url?scp=77955283399&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77955283399&partnerID=8YFLogxK

U2 - 10.1128/JB.00237-10

DO - 10.1128/JB.00237-10

M3 - Article

C2 - 20525824

AN - SCOPUS:77955283399

VL - 192

SP - 3883

EP - 3892

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 15

ER -