While powerful for studying pathogenic secreted effectors, structural biology has not been used to a large extent to study effector-host protein interactions. In this proposal, we use known and novel secreted effectors and an experimental pipeline that will provide broad insights into how a model pathogen. Salmonella, subverts host cell function.
Aim 1 simultaneously provides key functional insights into effector protein potency and function using broad set of assays, while at the same time serves as an important selection step to focus structural characterization by the PSI network on the most valuable targets.
Aim 2 provides valuable information about the host proteins and protein pathways, using cross-linking and proteomics, that each effector interacts with and provides the PSI network with a prioritized list of host protein targets for structural characterization.
Aim 3 interrogates possible specific protein-protein interactions and provides a selection of validated protein-protein interactions and possibly ligands for structural characterization by PSI network. Ultimately, structure-function studies of individual secreted effectors are invaluable to the host-pathogen research community. However, the insights into host-pathogen biology gained from this large parallel characterization effort with multiple effectors will advance understanding at a more complete systems level.
Advanced mass spectrometry methods with structure determinations will allow for improved understanding of the interactions between human host proteins and pathogens;this research may lead to new therapies for infectious diseases. The specific pathogen model to be studied here is Salmonella, a pathogen that has been the cause of a number of significant food-bourne outbreaks in recent years.
|Wu, Jikang; Sabag-Daigle, Anice; Borton, Mikayla A et al. (2018) Salmonella-Mediated Inflammation Eliminates Competitors for Fructose-Asparagine in the Gut. Infect Immun 86:|
|Merkley, Eric D; Sego, Landon H; Lin, Andy et al. (2017) Protein abundances can distinguish between naturally-occurring and laboratory strains of Yersinia pestis, the causative agent of plague. PLoS One 12:e0183478|
|Waters, Elaine M; Rudkin, Justine K; Coughlan, Simone et al. (2017) Redeploying ?-Lactam Antibiotics as a Novel Antivirulence Strategy for the Treatment of Methicillin-Resistant Staphylococcus aureus Infections. J Infect Dis 215:80-87|
|Lama, Amrita; Drennan, Samuel L; Johnson, Rudd C et al. (2017) Identification of Conserved ABC Importers Necessary for Intracellular Survival of Legionella pneumophila in Multiple Hosts. Front Cell Infect Microbiol 7:485|
|Negretti, Nicholas M; Gourley, Christopher R; Clair, Geremy et al. (2017) The food-borne pathogen Campylobacter jejuni responds to the bile salt deoxycholate with countermeasures to reactive oxygen species. Sci Rep 7:15455|
|Sontag, Ryan L; Nakayasu, Ernesto S; Brown, Roslyn N et al. (2016) Identification of Novel Host Interactors of Effectors Secreted by Salmonella and Citrobacter. mSystems 1:|
|Nakayasu, Ernesto S; Sydor, Michael A; Brown, Roslyn N et al. (2015) Identification of Salmonella Typhimurium Deubiquitinase SseL Substrates by Immunoaffinity Enrichment and Quantitative Proteomic Analysis. J Proteome Res 14:4029-38|
|Sontag, Ryan L; Mihai, Cosmin; Orr, Galya et al. (2015) Electroporation of functional bacterial effectors into mammalian cells. J Vis Exp :52296|
|Elfenbein, Johanna R; Knodler, Leigh A; Nakayasu, Ernesto S et al. (2015) Multicopy Single-Stranded DNA Directs Intestinal Colonization of Enteric Pathogens. PLoS Genet 11:e1005472|
|Li, Jie; Overall, Christopher C; Nakayasu, Ernesto S et al. (2015) Analysis of the Salmonella regulatory network suggests involvement of SsrB and H-NS in ?(E)-regulated SPI-2 gene expression. Front Microbiol 6:27|
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