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Kinase suppressor of Ras-1 protects against pulmonary Pseudomonas aeruginosa infections

Nature Medicine volume 17, pages 341–346 (2011)Cite this article

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Abstract

Pseudomonas aeruginosa is a Gram-negative pathogen that causes severe infections in immunocompromised individuals and individuals with cystic fibrosis or chronic obstructive pulmonary disease (COPD). Here we show that kinase suppressor of Ras-1 (Ksr1)-deficient mice are highly susceptible to pulmonary P. aeruginosa infection accompanied by uncontrolled pulmonary cytokine release, sepsis and death, whereas wild-type mice clear the infection. Ksr1 recruits and assembles inducible nitric oxide (NO) synthase (iNOS) and heat shock protein-90 (Hsp90) to enhance iNOS activity and to release NO upon infection. Ksr1 deficiency prevents lung alveolar macrophages and neutrophils from activating iNOS, producing NO and killing bacteria. Restoring NO production restores the bactericidal capability of Ksr1-deficient lung alveolar macrophages and neutrophils and rescues Ksr1-deficient mice from P. aeruginosa infection. Our findings suggest that Ksr1 functions as a previously unknown scaffold that enhances iNOS activity and is therefore crucial for the pulmonary response to P. aeruginosa infections.

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Figure 1: Ksr1 deficiency results in hypersusceptibility to pulmonary P. aeruginosa infection.
Figure 2: Decreased bacterial-killing capability in Ksr1-deficient alveolar macrophages.
Figure 3: Reduction of P. aeruginosa–induced NO production and peroxynitrite formation in Ksr1-deficient alveolar macrophages.
Figure 4: Ksr1 anchors and enhances the activity of iNOS.
Figure 5: Ksr1 assembles Hsp90 and iNOS to activate iNOS.
Figure 6: NO is crucial for the elimination of pulmonary P. aeruginosa.

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Acknowledgements

We thank R. Kolesnick (Memorial Sloan-Kettering Cancer Center) for providing us with Ksr1-deficient mice. This study was supported by Deutsche Forschungsgemeinschaft grants Gu 335/13-3 and Gu 335/16-2 to E.G.

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Author notes

  1. Yang Zhang and Xiang Li: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology, University of Duisburg-Essen, Essen, Germany

    Yang Zhang, Xiang Li, Alexander Carpinteiro, Matthias Soddemann & Erich Gulbins

  2. Department of Cell and Developmental Biology, Vanderbilt University School of Medicine and Monroe Carell Jr. Children's Hospital at Vanderbilt, Nashville, Tennessee, USA

    Jeremy A Goettel

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  2. Xiang Li
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Contributions

Y.Z. performed the internalization assays, confocal and fluorescence microscopy, immunoprecipitation studies, flow cytometry analysis and protein overexpression and iNOS activity assays. X.L. conducted the mouse infection experiments, determined cytokine abundance in the lung and performed the bacterial-killing assay and immunoblot analysis. A.C. performed bone marrow transplantation and determined the neutrophil count. M.S. characterized the genotypes of Ksr1-deficient mice. J.A.G. cloned recombinant Ksr proteins. Y.Z., X.L. and E.G. designed the experiments and wrote the manuscript.

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Correspondence to Erich Gulbins.

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The authors declare no competing financial interests.

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Supplementary Notes 1–3, Supplementary Figures 1–6 and Supplementary Methods (PDF 463 kb)

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Zhang, Y., Li, X., Carpinteiro, A. et al. Kinase suppressor of Ras-1 protects against pulmonary Pseudomonas aeruginosa infections. Nat Med 17, 341–346 (2011). https://doi.org/10.1038/nm.2296

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