title: Comparative phosphoproteomics reveals components of host cell invasion and post-transcriptional regulation during Francisella infection.
abstract: Francisella tularensis is a facultative intracellular bacterium that causes the deadly disease tularemia. Most evidence suggests that Francisella is not well recognized by the innate immune system that normally leads to cytokine expression and cell death. In previous work, we identified new bacterial factors that were hyper-cytotoxic to macrophages. Four of the identified hyper-cytotoxic strains (lpcC, manB, manC, and kdtA) had an impaired lipopolysaccharide (LPS) synthesis and produced an exposed lipid A lacking the O-antigen. These mutants were not only hyper-cytotoxic but also were phagocytosed at much higher rates compared with the wild type parent strain. To elucidate the cellular signaling underlying this enhanced phagocytosis and cell death, we performed a large-scale comparative phosphoproteomic analysis of cells infected with wild-type and delta-lpcC F. novicida. Our data suggest that not only actin but also intermediate filaments and microtubules are important for F. novicida entry into the host cells. In addition, we observed differential phosphorylation of tristetraprolin, a key component of the mRNA-degrading machinery that controls the expression of a variety of genes including many cytokines. Infection with the delta-lpcC mutant induced the hyper-phosphorylation and inhibition of tristetraprolin, leading to the production of cytokines such as IL-1beta and TNF-alpha that may kill the host cells by triggering apoptosis. Together, our data provide new insights for Francisella invasion and a post-transcriptional mechanism that prevents the expression of host immune response factors that control infection by this pathogen.
authors: Biological Science Division, Pacific Northwest National Laboratory, Richland, Washington;
authors: Ernesto S
authors: ES
authors: Nakayasu
authors: Rebecca
authors: R
authors: Tempel
authors: Xiaolu A
authors: XA
authors: Cambronne
authors: Vladislav A
authors: VA
authors: Petyuk
authors: Marcus B
authors: MB
authors: Jones
authors: Marina A
authors: MA
authors: Gritsenko
authors: Matthew E
authors: ME
authors: Monroe
authors: Feng
authors: F
authors: Yang
authors: Richard D
authors: RD
authors: Smith
authors: Joshua N
authors: JN
authors: Adkins
authors: Fred
authors: F
authors: Heffron
mesh_terms: D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005603:Francisella / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D005798:Genes, Bacterial; D016905:Gram-Negative Bacterial Infections / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D054884:Host-Pathogen Interactions / Q000235:genetics / Q000276:immunology; D007113:Immunity, Innate; D008264:Macrophages / Q000276:immunology / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D060066:Protein Interaction Maps; D040901:Proteomics; D012313:RNA / Q000235:genetics / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D014774:Virulence / Q000235:genetics / Q000502:physiology
chemical_list: D016207:Cytokines; D010750:Phosphoproteins; D012313:RNA
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