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 Article · Fri Sep 01 04:00:00 EDT 2006 · Journal of Biological Chemistry, 281:29131-29140

DOI:https://doi.org/10.1074/jbc.M604640200· OSTI ID:892897

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

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.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 892897
• Report Number(s): PNNL-SA-51416; 11491
• Journal Information: Journal of Biological Chemistry, 281:29131-29140, Journal Name: Journal of Biological Chemistry, 281:29131-29140 Vol. 281; ISSN JBCHA3; ISSN 0021-9258
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
99 GENERAL AND MISCELLANEOUS
ABUNDANCE
BIOSYNTHESIS
CELL WALL
Environmental Molecular Sciences Laboratory
FUNCTIONALS
GENES
MACROPHAGES
MODIFICATIONS
PATHOGENESIS
PROTEINS
SALMONELLA
SPECTROSCOPY
VIRULENCE