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Intracellular bacterial growth is controlled by a kinase network around PKB/AKT1

Nature volume 450, pages 725–730 (2007)Cite this article

Abstract

With the emergence of multidrug resistant (MDR) bacteria, it is imperative to develop new intervention strategies. Current antibiotics typically target pathogen rather than host-specific biochemical pathways1. Here we have developed kinase inhibitors that prevent intracellular growth of unrelated pathogens such as Salmonella typhimurium and Mycobacterium tuberculosis. An RNA interference screen of the human kinome using automated microscopy revealed several host kinases capable of inhibiting intracellular growth of S. typhimurium. The kinases identified clustered in one network around AKT1 (also known as PKB). Inhibitors of AKT1 prevent intracellular growth of various bacteria including MDR-M. tuberculosis. AKT1 is activated by the S. typhimurium effector SopB, which promotes intracellular survival by controlling actin dynamics through PAK4, and phagosome–lysosome fusion through the AS160 (also known as TBC1D4)–RAB14 pathway. AKT1 inhibitors counteract the bacterial manipulation of host signalling processes, thus controlling intracellular growth of bacteria. By using a reciprocal chemical genetics approach, we identified kinase inhibitors with antibiotic properties and their host targets, and we determined host signalling networks that are activated by intracellular bacteria for survival.

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Figure 1: The protein-kinase-A inhibitor H-89 inhibits host kinases that control intracellular Salmonella typhimurium growth.
Figure 2: Chemical profiling for antibiotic activity of kinase inhibitors.
Figure 3: Identification of host kinases controlling intracellular growth of S. typhimurium.
Figure 4: Identification of host kinase targets for the chemical compounds inhibiting S. typhimurium and (MDR-) M. tuberculosis.
Figure 5: Mechanisms of AKT1 control of intracellular S. typhimurium infections.

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Acknowledgements

This work was supported by a program grant from N.W.O. and the Dutch Cancer Society KWF to J.N. and T.H.M.O., and an EEC network grant (Microban). We thank J. Sung for the histochemical analyses of mouse tissues, W. Mooi and W. Zwart for kinase-compound modelling, S. Commandeur and K. Walburg for support in the chemical-profiling experiments, D. Holden for SopB-deficient S. typhimurium, L. Wilson for help with culturing M. tuberculosis strains, H. v. d. Elst for assistance in purification and LCMS analysis of the chemical inhibitors, K. Kremer for providing M. tuberculosis clinical isolates, K. Nealson and S. Vesterlund for Lux-S. typhimurium, C. Wells and A. Ridley for haemagglutinin-tagged PAK4 contructs, and G. Lienhard for AS160 constructs.

Author Contributions C.K. performed the kinase assays (with J.N.) and experiments with RAB14, AS160 (with M.K.) and PAK4. M.M. and C.K. performed the shRNAi screen with a library made by R.L.B. and obtained from D.A.E. C.K. performed the siRNA screening, automated microscopy and data analyses, supported by the NKI robotics facility (R.L.B., D.E.). Subcloning and sequencing was by L.J. A.T., R.v.d.N. and A.P. synthesized the kinase inhibitors under the supervision of H.O. and G.v.d.M. N.D.L.S., S.J.F.v.d.E. and A.G. performed in vivo and chemical-profiling experiments (supervised by T.H.M.O.). J.N. supervised the study.

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Authors and Affiliations

  1. Division of Tumor Biology, and, Amsterdam

    Coenraad Kuijl, Marije Marsman, Lennert Janssen, Mirjam Ketema & Jacques Neefjes

  2. Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands, Amsterdam

    David A. Egan & Roderick L. Beijersbergen

  3. Department of Immunohematology and Blood Transfusion and Department of Infectious Diseases, Leiden Medical University Centre, Albinusdreef 2, 2333 ZA Leiden, The Netherlands, Leiden

    Nigel D. L. Savage, Susan J. F. van den Eeden, Annemieke Geluk & Tom H. M. Ottenhoff

  4. Department of Bioorganic Chemistry, Leiden Institute of Chemistry, 2300 RA Leiden, The Netherlands

    Adriaan W. Tuin, Rian van den Nieuwendijk, Alex Poot, Gijs van der Marel & Hermen Overkleeft

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Correspondence to Jacques Neefjes.

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Kuijl, C., Savage, N., Marsman, M. et al. Intracellular bacterial growth is controlled by a kinase network around PKB/AKT1. Nature 450, 725–730 (2007). https://doi.org/10.1038/nature06345

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