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Title: 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

Abstract

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 (LC-MS)-based proteomic approach to detect the changes in STM protein abundances. 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 isolated STM cells, which are largely house-keeping 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 tomore » 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 STM cell wall, emphasizing their important roles in the colonization of macrophages by Salmonella.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
892897
Report Number(s):
PNNL-SA-51416
Journal ID: ISSN 0021-9258; JBCHA3; 11491; TRN: US200716%%583
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Journal of Biological Chemistry, 281:29131-29140
Additional Journal Information:
Journal Volume: 281; Journal ID: ISSN 0021-9258
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; ABUNDANCE; BIOSYNTHESIS; CELL WALL; FUNCTIONALS; GENES; MACROPHAGES; MODIFICATIONS; PATHOGENESIS; PROTEINS; SALMONELLA; SPECTROSCOPY; VIRULENCE; Environmental Molecular Sciences Laboratory

Citation Formats

Shi, Liang, Adkins, Joshua N, Coleman, James R, Schepmoes, Athena A, Dohnalkova, Alice, Mottaz, Heather M, Norbeck, Angela D, Purvine, Samuel O, Manes, Nathan P, Smallwood, Heather S, Wang, Haixing H, Forbes, John, Gros, Philippe, Uzzau, Sergio, Rodland, Karin D, Heffron, Fred, Smith, Richard D, and 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. United States: N. p., 2006. Web. doi:10.1074/jbc.M604640200.
Shi, Liang, Adkins, Joshua N, Coleman, James R, Schepmoes, Athena A, Dohnalkova, Alice, Mottaz, Heather M, Norbeck, Angela D, Purvine, Samuel O, Manes, Nathan P, Smallwood, Heather S, Wang, Haixing H, 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. United States. https://doi.org/10.1074/jbc.M604640200
Shi, Liang, Adkins, Joshua N, Coleman, James R, Schepmoes, Athena A, Dohnalkova, Alice, Mottaz, Heather M, Norbeck, Angela D, Purvine, Samuel O, Manes, Nathan P, Smallwood, Heather S, Wang, Haixing H, Forbes, John, Gros, Philippe, Uzzau, Sergio, Rodland, Karin D, Heffron, Fred, Smith, Richard D, and Squier, Thomas C. 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". United States. https://doi.org/10.1074/jbc.M604640200.
@article{osti_892897,
title = {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},
author = {Shi, Liang and Adkins, Joshua N and Coleman, James R and Schepmoes, Athena A and Dohnalkova, Alice and Mottaz, Heather M and Norbeck, Angela D and Purvine, Samuel O and Manes, Nathan P and Smallwood, Heather S and Wang, Haixing H and Forbes, John and Gros, Philippe and Uzzau, Sergio and Rodland, Karin D and Heffron, Fred and Smith, Richard D and Squier, Thomas C},
abstractNote = {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 (LC-MS)-based proteomic approach to detect the changes in STM protein abundances. 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 isolated STM cells, which are largely house-keeping 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 STM cell wall, emphasizing their important roles in the colonization of macrophages by Salmonella.},
doi = {10.1074/jbc.M604640200},
url = {https://www.osti.gov/biblio/892897}, journal = {Journal of Biological Chemistry, 281:29131-29140},
issn = {0021-9258},
number = ,
volume = 281,
place = {United States},
year = {Fri Sep 01 00:00:00 EDT 2006},
month = {Fri Sep 01 00:00:00 EDT 2006}
}