Proteomic analysis of Salmonella enterica serovar Typhimurium isolated from RAW 264.7 macrophages: Identification of a novel protein that contributes to the replication of serovar Typhimurium inside macrophages

Liang Shi, Joshua N. Adkins, James R. Coleman, Athena A. Schepmoes, Alice Dohnkova, Heather M. Mottaz, Angela D. Norbeck, Samuel O. Purvine, Nathan P. Manes, Heather S. Smallwood, Haixing Wang, John Forbes, Philippe Gros, Sergio Uzzau, Karin D. Rodland, Fred Heffron, Richard D. Smith, Thomas C. Squier

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

To evade host resistance mechanisms, Salmonella enterica serovar Typhimurium (STM), a facultative intracellular pathogen, must alter its proteome following macrophage infection. To identify new colonization and virulence factors that mediate STM pathogenesis, we have isolated STM cells from RAW 264.7 macrophages at various time points following infection and used a liquid chromatography-mass spectrometry-based proteomic approach to detect the changes in STM protein abundance. Because host resistance to STM infection is strongly modulated by the expression of a functional host-resistant regulator, i.e. natural resistance-associated macrophage protein 1 (Nramp1, also called Slc11a1),we have also examined the effects of Nramp1 activity on the changes of STM protein abundances. A total of 315 STM proteins have been identified from isolatedSTMcells, which are largely housekeeping proteins whose abundances remain relatively constant during the time course of infection. However, 39 STM proteins are strongly induced after infection, suggesting their involvement in modulating colonization and infection. Of the 39 induced proteins, 6 proteins are specifically modulated by Nramp1 activity, including STM3117, as well as STM3118-3119 whose time-dependent abundance changes were confirmed using Western blot analysis. Deletion of the gene encoding STM3117 resulted in a dramatic reduction in the ability of STM to colonize wild-type RAW 264.7 macrophages, demonstrating a critical involvement of STM3117 in promoting the replication of STM inside macrophages. The predicted function common for STM3117-3119 is biosynthesis and modification of the peptidoglycan layer of the STM cell wall.

Original languageEnglish (US)
Pages (from-to)29131-29140
Number of pages10
JournalJournal of Biological Chemistry
Volume281
Issue number39
DOIs
StatePublished - Sep 29 2006

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Salmonella
Salmonella enterica
Macrophages
Proteomics
Proteins
Infection
Gene encoding
Peptidoglycan
Biosynthesis
Liquid chromatography
Virulence Factors
Pathogens
Proteome
Serogroup
Mass spectrometry
Housekeeping
Cells
Gene Deletion
Liquid Chromatography
Cell Wall

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Proteomic analysis of Salmonella enterica serovar Typhimurium isolated from RAW 264.7 macrophages : Identification of a novel protein that contributes to the replication of serovar Typhimurium inside macrophages. / Shi, Liang; Adkins, Joshua N.; Coleman, James R.; Schepmoes, Athena A.; Dohnkova, Alice; Mottaz, Heather M.; Norbeck, Angela D.; Purvine, Samuel O.; Manes, Nathan P.; Smallwood, Heather S.; Wang, Haixing; Forbes, John; Gros, Philippe; Uzzau, Sergio; Rodland, Karin D.; Heffron, Fred; Smith, Richard D.; Squier, Thomas C.

In: Journal of Biological Chemistry, Vol. 281, No. 39, 29.09.2006, p. 29131-29140.

Research output: Contribution to journalArticle

Shi, L, Adkins, JN, Coleman, JR, Schepmoes, AA, Dohnkova, A, Mottaz, HM, Norbeck, AD, Purvine, SO, Manes, NP, Smallwood, HS, Wang, H, Forbes, J, Gros, P, Uzzau, S, Rodland, KD, Heffron, F, Smith, RD & Squier, TC 2006, 'Proteomic analysis of Salmonella enterica serovar Typhimurium isolated from RAW 264.7 macrophages: Identification of a novel protein that contributes to the replication of serovar Typhimurium inside macrophages', Journal of Biological Chemistry, vol. 281, no. 39, pp. 29131-29140. https://doi.org/10.1074/jbc.M604640200
Shi, Liang ; Adkins, Joshua N. ; Coleman, James R. ; Schepmoes, Athena A. ; Dohnkova, Alice ; Mottaz, Heather M. ; Norbeck, Angela D. ; Purvine, Samuel O. ; Manes, Nathan P. ; Smallwood, Heather S. ; Wang, Haixing ; Forbes, John ; Gros, Philippe ; Uzzau, Sergio ; Rodland, Karin D. ; Heffron, Fred ; Smith, Richard D. ; Squier, Thomas C. / Proteomic analysis of Salmonella enterica serovar Typhimurium isolated from RAW 264.7 macrophages : Identification of a novel protein that contributes to the replication of serovar Typhimurium inside macrophages. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 39. pp. 29131-29140.
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abstract = "To evade host resistance mechanisms, Salmonella enterica serovar Typhimurium (STM), a facultative intracellular pathogen, must alter its proteome following macrophage infection. To identify new colonization and virulence factors that mediate STM pathogenesis, we have isolated STM cells from RAW 264.7 macrophages at various time points following infection and used a liquid chromatography-mass spectrometry-based proteomic approach to detect the changes in STM protein abundance. Because host resistance to STM infection is strongly modulated by the expression of a functional host-resistant regulator, i.e. natural resistance-associated macrophage protein 1 (Nramp1, also called Slc11a1),we have also examined the effects of Nramp1 activity on the changes of STM protein abundances. A total of 315 STM proteins have been identified from isolatedSTMcells, which are largely housekeeping proteins whose abundances remain relatively constant during the time course of infection. However, 39 STM proteins are strongly induced after infection, suggesting their involvement in modulating colonization and infection. Of the 39 induced proteins, 6 proteins are specifically modulated by Nramp1 activity, including STM3117, as well as STM3118-3119 whose time-dependent abundance changes were confirmed using Western blot analysis. Deletion of the gene encoding STM3117 resulted in a dramatic reduction in the ability of STM to colonize wild-type RAW 264.7 macrophages, demonstrating a critical involvement of STM3117 in promoting the replication of STM inside macrophages. The predicted function common for STM3117-3119 is biosynthesis and modification of the peptidoglycan layer of the STM cell wall.",
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AU - Shi, Liang

AU - Adkins, Joshua N.

AU - Coleman, James R.

AU - Schepmoes, Athena A.

AU - Dohnkova, Alice

AU - Mottaz, Heather M.

AU - Norbeck, Angela D.

AU - Purvine, Samuel O.

AU - Manes, Nathan P.

AU - Smallwood, Heather S.

AU - Wang, Haixing

AU - Forbes, John

AU - Gros, Philippe

AU - Uzzau, Sergio

AU - Rodland, Karin D.

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AU - Smith, Richard D.

AU - Squier, Thomas C.

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N2 - To evade host resistance mechanisms, Salmonella enterica serovar Typhimurium (STM), a facultative intracellular pathogen, must alter its proteome following macrophage infection. To identify new colonization and virulence factors that mediate STM pathogenesis, we have isolated STM cells from RAW 264.7 macrophages at various time points following infection and used a liquid chromatography-mass spectrometry-based proteomic approach to detect the changes in STM protein abundance. Because host resistance to STM infection is strongly modulated by the expression of a functional host-resistant regulator, i.e. natural resistance-associated macrophage protein 1 (Nramp1, also called Slc11a1),we have also examined the effects of Nramp1 activity on the changes of STM protein abundances. A total of 315 STM proteins have been identified from isolatedSTMcells, which are largely housekeeping proteins whose abundances remain relatively constant during the time course of infection. However, 39 STM proteins are strongly induced after infection, suggesting their involvement in modulating colonization and infection. Of the 39 induced proteins, 6 proteins are specifically modulated by Nramp1 activity, including STM3117, as well as STM3118-3119 whose time-dependent abundance changes were confirmed using Western blot analysis. Deletion of the gene encoding STM3117 resulted in a dramatic reduction in the ability of STM to colonize wild-type RAW 264.7 macrophages, demonstrating a critical involvement of STM3117 in promoting the replication of STM inside macrophages. The predicted function common for STM3117-3119 is biosynthesis and modification of the peptidoglycan layer of the STM cell wall.

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