TY - JOUR
T1 - Identification of novel host interactors of effectors secreted by Salmonella and Citrobacter
AU - Sontag, Ryan L.
AU - Nakayasu, Ernesto S.
AU - Brown, Roslyn N.
AU - Niemann, George S.
AU - Sydor, Michael A.
AU - Sanchez, Octavio
AU - Ansong, Charles
AU - Lu, Shao Yeh
AU - Choi, Hyungwon
AU - Valleau, Dylan
AU - Weitz, Karl K.
AU - Savchenko, Alexei
AU - Cambronne, Eric D.
AU - Adkins, Joshua N.
N1 - Funding Information:
This work, including the efforts of Ryan L. Sontag, Ernesto S. Nakayasu, Roslyn N. Brown, George S. Niemann, Michael A. Sydor, Octavio Sanchez, Charles Ansong, Shao-Yeh Lu, Hyungwon Choi, Karl K. Weitz, Eric D. Cambronne, and Joshua N. Adkins, was funded by HHS | National Institutes of Health (NIH) (GM094623). This work, including the efforts of Ernesto S. Nakayasu, Charles Ansong, and Joshua N. Adkins, was funded by HHS | National Institutes of Health (NIH) (GM103493-10). This work, including the efforts of Dylan Valleau and Alexei Savchenko, was funded by HHS | National Institutes of Health (NIH) (GM094585).
Funding Information:
This research was funded in part by grants from the National Institutes of Health (grants GM094585, GM094623, and P41 GM103493-10). The work was partly performed in the Environmental Molecular Sciences Laboratory (EMSL), a DOE-BER national scientific user facility at Pacific Northwest National Laboratory (PNNL). PNNL is a multiprogram national laboratory operated by Battelle Memorial Institute for the DOE under contract DE-AC05-76RLO 1830.
Publisher Copyright:
Copyright © 2016 Sontag et al.
PY - 2016/7/1
Y1 - 2016/7/1
N2 - Many pathogenic bacteria of the family Enterobacteriaceae use type III secretion systems to inject virulence proteins, termed "effectors," into the host cell cytosol. Although host-cellular activities of several effectors have been demonstrated, the function and host-targeted pathways of most of the effectors identified to date are largely undetermined. To gain insight into host proteins targeted by bacterial effectors, we performed coaffinity purification of host proteins from cell lysates using recombinant effectors from the Enterobacteriaceae intracellular pathogens Salmonella enterica serovar Typhimurium and Citrobacter rodentium. We identified 54 high-confidence host interactors for the Salmonella effectors GogA, GtgA, GtgE, SpvC, SrfH, SseL, SspH1, and SssB collectively and 21 interactors for the Citrobacter effectors EspT, NleA, NleG1, and NleK. We biochemically validated the interaction between the SrfH Salmonella protein and the extracellular signal-regulated kinase 2 (ERK2) host protein kinase, which revealed a role for this effector in regulating phosphorylation levels of this enzyme, which plays a central role in signal transduction.
AB - Many pathogenic bacteria of the family Enterobacteriaceae use type III secretion systems to inject virulence proteins, termed "effectors," into the host cell cytosol. Although host-cellular activities of several effectors have been demonstrated, the function and host-targeted pathways of most of the effectors identified to date are largely undetermined. To gain insight into host proteins targeted by bacterial effectors, we performed coaffinity purification of host proteins from cell lysates using recombinant effectors from the Enterobacteriaceae intracellular pathogens Salmonella enterica serovar Typhimurium and Citrobacter rodentium. We identified 54 high-confidence host interactors for the Salmonella effectors GogA, GtgA, GtgE, SpvC, SrfH, SseL, SspH1, and SssB collectively and 21 interactors for the Citrobacter effectors EspT, NleA, NleG1, and NleK. We biochemically validated the interaction between the SrfH Salmonella protein and the extracellular signal-regulated kinase 2 (ERK2) host protein kinase, which revealed a role for this effector in regulating phosphorylation levels of this enzyme, which plays a central role in signal transduction.
KW - Affinity purification
KW - Effectors
KW - Mass spectrometry
KW - Pathogenic bacteria
KW - Protein-protein interactions
KW - Type III secretion system
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U2 - 10.1128/mSystems.00032-15
DO - 10.1128/mSystems.00032-15
M3 - Article
AN - SCOPUS:85021049950
SN - 2379-5077
VL - 1
JO - mSystems
JF - mSystems
IS - 4
M1 - e00032
ER -