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 funcfion.
Aim 1 simultaneously provides key functional insights into effector protein potency and funcfion using broad set of assays, while at the same fime 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 characterizafion by PSI network. Ultimately, structure-funcfion 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 characterizafion 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.
|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:|
|Sontag, Ryan L; Mihai, Cosmin; Orr, Galya et al. (2015) Electroporation of functional bacterial effectors into mammalian cells. J Vis Exp :52296|
|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|
|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 Ïƒ(E)-regulated SPI-2 gene expression. Front Microbiol 6:27|
|Li, Jie; Nakayasu, Ernesto S; Overall, Christopher C et al. (2015) Global analysis of Salmonella alternative sigma factor E on protein translation. J Proteome Res 14:1716-26|
|Eletsky, Alexander; Michalska, Karolina; Houliston, Scott et al. (2014) Structural and functional characterization of DUF1471 domains of Salmonella proteins SrfN, YdgH/SssB, and YahO. PLoS One 9:e101787|
|Lu, Jun; Wu, Ruiying; Adkins, Joshua N et al. (2014) Crystal structures of the F and pSLT plasmid TraJ N-terminal regions reveal similar homodimeric PAS folds with functional interchangeability. Biochemistry 53:5810-9|
|Merkley, Eric D; Rysavy, Steven; Kahraman, Abdullah et al. (2014) Distance restraints from crosslinking mass spectrometry: mining a molecular dynamics simulation database to evaluate lysine-lysine distances. Protein Sci 23:747-59|
|Nakayasu, Ernesto S; Brown, Roslyn N; Ansong, Charles et al. (2013) Multi-omic data integration links deleted in breast cancer 1 (DBC1) degradation to chromatin remodeling in inflammatory response. Mol Cell Proteomics 12:2136-47|
Showing the most recent 10 out of 18 publications