title: Bacterial virulence proteins as tools to rewire kinase pathways in yeast and immune cells.
abstract: Bacterial pathogens have evolved specific effector proteins that, by interfacing with host kinase signalling pathways, provide a mechanism to evade immune responses during infection. Although these effectors contribute to pathogen virulence, we realized that they might also serve as valuable synthetic biology reagents for engineering cellular behaviour. Here we exploit two effector proteins, the Shigella flexneri OspF protein and Yersinia pestis YopH protein, to rewire kinase-mediated responses systematically both in yeast and mammalian immune cells. Bacterial effector proteins can be directed to inhibit specific mitogen-activated protein kinase pathways selectively in yeast by artificially targeting them to pathway-specific complexes. Moreover, we show that unique properties of the effectors generate new pathway behaviours: OspF, which irreversibly inactivates mitogen-activated protein kinases, was used to construct a synthetic feedback circuit that shows novel frequency-dependent input filtering. Finally, we show that effectors can be used in T cells, either as feedback modulators to tune the T-cell response amplitude precisely, or as an inducible pause switch that can temporarily disable T-cell activation. These studies demonstrate how pathogens could provide a rich toolkit of parts to engineer cells for therapeutic or biotechnological applications.
authors: Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California 94158, USA.
authors: Ping
authors: P
authors: Wei
authors: Wilson W
authors: WW
authors: Wong
authors: Jason S
authors: JS
authors: Park
authors: Ethan E
authors: EE
authors: Corcoran
authors: Sergio G
authors: SG
authors: Peisajovich
authors: James J
authors: JJ
authors: Onuffer
authors: Arthur
authors: A
authors: Weiss
authors: Wendell A
authors: WA
authors: Lim
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D025461:Feedback, Physiological; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D007376:Interleukin-2 / Q000276:immunology; D019169:Jurkat Cells; D008213:Lymphocyte Activation / Q000235:genetics; D020935:MAP Kinase Signaling System; D009994:Osmolar Concentration; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D012763:Shigella flexneri / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D013601:T-Lymphocytes / Q000166:cytology / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*; D015010:Yersinia pestis / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D007376:Interleukin-2; D037521:Virulence Factors; D017027:Protein Tyrosine Phosphatases; C067571:yopH protein, Yersinia