The NEAT domain-containing proteins of Clostridium perfringens bind heme

Jocelyn M. Choo, Jackie K. Cheung, Jessica A. Wisniewski, David L. Steer, Dieter M. Bulach, Thomas J. Hiscox, Anjana Chakravorty, A. Ian Smith, David A. Gell, Julian I. Rood, Milena M. Awad

Research output: Contribution to journalArticleResearchpeer-review

5 Citations (Scopus)

Abstract

The ability of a pathogenic bacterium to scavenge iron from its host is important for its growth and survival during an infection. Our studies on Cperfringens gas gangrene strain JIR325, a derivative of strain 13, showed that it is capable of utilizing both human hemoglobin and ferric chloride, but not human holo-transferrin, as an iron source for in vitro growth. Analysis of the Cperfringens strain 13 genome sequence identified a putative heme acquisition system encoded by an iron-regulated surface gene region that we have named the Cht (Clostridium perfringensheme transport) locus. This locus comprises eight genes that are co-transcribed and includes genes that encode NEAT domain-containing proteins (ChtD and ChtE) and a putative sortase (Srt). The ChtD, ChtE and Srt proteins were shown to be expressed in JIR325 cells grown under iron-limited conditions and were localized to the cell envelope. Moreover, the NEAT proteins, ChtD and ChtE, were found to bind heme. Both chtDE and srt mutants were constructed, but these mutants were not defective in hemoglobin or ferric chloride utilization. They were, however, attenuated for virulence when tested in a mouse myonecrosis model, although the virulence phenotype could not be restored via complementation and, as is common with such systems, secondary mutations were identified in these strains. In summary, this study provides evidence for the functional redundancies that occur in the heme transport pathways of this life threatening pathogen.

Original languageEnglish
Article number0162981
Number of pages26
JournalPLoS ONE
Volume11
Issue number9
DOIs
Publication statusPublished - 16 Sep 2016

Keywords

  • heme
  • clostridium perfringens
  • genetic loci
  • staphylococcus aureus
  • polymerase chain reaction
  • bacillus anthracis
  • plasmid construction
  • recombinant proteins

Cite this

Choo, Jocelyn M. ; Cheung, Jackie K. ; Wisniewski, Jessica A. ; Steer, David L. ; Bulach, Dieter M. ; Hiscox, Thomas J. ; Chakravorty, Anjana ; Smith, A. Ian ; Gell, David A. ; Rood, Julian I. ; Awad, Milena M. / The NEAT domain-containing proteins of Clostridium perfringens bind heme. In: PLoS ONE. 2016 ; Vol. 11, No. 9.
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abstract = "The ability of a pathogenic bacterium to scavenge iron from its host is important for its growth and survival during an infection. Our studies on C. perfringens gas gangrene strain JIR325, a derivative of strain 13, showed that it is capable of utilizing both human hemoglobin and ferric chloride, but not human holo-transferrin, as an iron source for in vitro growth. Analysis of the C. perfringens strain 13 genome sequence identified a putative heme acquisition system encoded by an iron-regulated surface gene region that we have named the Cht (Clostridium perfringensheme transport) locus. This locus comprises eight genes that are co-transcribed and includes genes that encode NEAT domain-containing proteins (ChtD and ChtE) and a putative sortase (Srt). The ChtD, ChtE and Srt proteins were shown to be expressed in JIR325 cells grown under iron-limited conditions and were localized to the cell envelope. Moreover, the NEAT proteins, ChtD and ChtE, were found to bind heme. Both chtDE and srt mutants were constructed, but these mutants were not defective in hemoglobin or ferric chloride utilization. They were, however, attenuated for virulence when tested in a mouse myonecrosis model, although the virulence phenotype could not be restored via complementation and, as is common with such systems, secondary mutations were identified in these strains. In summary, this study provides evidence for the functional redundancies that occur in the heme transport pathways of this life threatening pathogen.",
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The NEAT domain-containing proteins of Clostridium perfringens bind heme. / Choo, Jocelyn M.; Cheung, Jackie K.; Wisniewski, Jessica A.; Steer, David L.; Bulach, Dieter M.; Hiscox, Thomas J.; Chakravorty, Anjana; Smith, A. Ian; Gell, David A.; Rood, Julian I.; Awad, Milena M.

In: PLoS ONE, Vol. 11, No. 9, 0162981, 16.09.2016.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - The NEAT domain-containing proteins of Clostridium perfringens bind heme

AU - Choo, Jocelyn M.

AU - Cheung, Jackie K.

AU - Wisniewski, Jessica A.

AU - Steer, David L.

AU - Bulach, Dieter M.

AU - Hiscox, Thomas J.

AU - Chakravorty, Anjana

AU - Smith, A. Ian

AU - Gell, David A.

AU - Rood, Julian I.

AU - Awad, Milena M.

PY - 2016/9/16

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N2 - The ability of a pathogenic bacterium to scavenge iron from its host is important for its growth and survival during an infection. Our studies on C. perfringens gas gangrene strain JIR325, a derivative of strain 13, showed that it is capable of utilizing both human hemoglobin and ferric chloride, but not human holo-transferrin, as an iron source for in vitro growth. Analysis of the C. perfringens strain 13 genome sequence identified a putative heme acquisition system encoded by an iron-regulated surface gene region that we have named the Cht (Clostridium perfringensheme transport) locus. This locus comprises eight genes that are co-transcribed and includes genes that encode NEAT domain-containing proteins (ChtD and ChtE) and a putative sortase (Srt). The ChtD, ChtE and Srt proteins were shown to be expressed in JIR325 cells grown under iron-limited conditions and were localized to the cell envelope. Moreover, the NEAT proteins, ChtD and ChtE, were found to bind heme. Both chtDE and srt mutants were constructed, but these mutants were not defective in hemoglobin or ferric chloride utilization. They were, however, attenuated for virulence when tested in a mouse myonecrosis model, although the virulence phenotype could not be restored via complementation and, as is common with such systems, secondary mutations were identified in these strains. In summary, this study provides evidence for the functional redundancies that occur in the heme transport pathways of this life threatening pathogen.

AB - The ability of a pathogenic bacterium to scavenge iron from its host is important for its growth and survival during an infection. Our studies on C. perfringens gas gangrene strain JIR325, a derivative of strain 13, showed that it is capable of utilizing both human hemoglobin and ferric chloride, but not human holo-transferrin, as an iron source for in vitro growth. Analysis of the C. perfringens strain 13 genome sequence identified a putative heme acquisition system encoded by an iron-regulated surface gene region that we have named the Cht (Clostridium perfringensheme transport) locus. This locus comprises eight genes that are co-transcribed and includes genes that encode NEAT domain-containing proteins (ChtD and ChtE) and a putative sortase (Srt). The ChtD, ChtE and Srt proteins were shown to be expressed in JIR325 cells grown under iron-limited conditions and were localized to the cell envelope. Moreover, the NEAT proteins, ChtD and ChtE, were found to bind heme. Both chtDE and srt mutants were constructed, but these mutants were not defective in hemoglobin or ferric chloride utilization. They were, however, attenuated for virulence when tested in a mouse myonecrosis model, although the virulence phenotype could not be restored via complementation and, as is common with such systems, secondary mutations were identified in these strains. In summary, this study provides evidence for the functional redundancies that occur in the heme transport pathways of this life threatening pathogen.

KW - heme

KW - clostridium perfringens

KW - genetic loci

KW - staphylococcus aureus

KW - polymerase chain reaction

KW - bacillus anthracis

KW - plasmid construction

KW - recombinant proteins

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