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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 infection1,2. 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 protein3 and Yersinia pestis YopH protein4, 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 kinases4, 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.

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Figure 1: Bacterial effector OspF can block selective MAPK pathways in yeast.
Figure 2: Tuning-frequency-dependent response of yeast osmolarity pathway using synthetic OspF feedback loop.
Figure 3: OspF can be used to control T-cell activation amplitude and duration precisely in Jurkat T cells.
Figure 4: OspF can be used as a synthetic pause switch to control human primary CD4 + T-cell activation.

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Acknowledgements

We thank K. Orth for the YopH plasmid; K. McNally and S. Neou for tissue culture support; H. El-Samad, C. Voigt, C. Tang and the Lim laboratory for discussions. We acknowledge the 2007 UCSF iGEM team (M. Chen, E. Chou, J. Huang, L. Jann, E. Meltzer, A. Ng and R. Ovadia) for their initial work on bacterial effectors in yeast. This work was supported by American Cancer Society fellowship PF-09-137-01-TBE (W.W.W), a Li Foundation Fellowship (P.W.), a California Institute for Regenerative Medicine fellowship (grant number TG2-01153) (J.S.P.), National Institutes of Health grants PN2EY016546, RO1GM055040, RO1GM062583 and P50GM081879 (W.A.L.), the NSF Synthetic Biology and Engineering Research Center (W.A.L.), the Packard Foundation (W.A.L.) and the Howard Hughes Medical Institute (A.W. and W.A.L.).

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Contributions

P.W., S.G.P. and W.A.L. initiated the project in yeast. W.W.W., E.E.C., A.W. and W.A.L. initiated the project in T cells. P.W., W.W.W. and W.A.L. wrote the paper. P.W. and S.G.P. planned and performed the experiments in yeast. W.W.W., P.W., E.E.C., J.J.O. and J.S.P. planned and performed the experiment in T cells.

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Correspondence to Wendell A. Lim.

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Wei, P., Wong, W., Park, J. et al. Bacterial virulence proteins as tools to rewire kinase pathways in yeast and immune cells. Nature 488, 384–388 (2012). https://doi.org/10.1038/nature11259

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