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 - 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 -