Salmonella infection impacts host proteome thermal stability

European Journal of Cell Biology, vol. 103, pp. 151448

Abstract

Intracellular bacterial pathogens hijack the protein machinery of infected host cells to evade their defenses and cultivate a favorable intracellular niche. The intracellular pathogen Salmonella enterica subsp. Typhimurium (STm) achieves this by injecting a cocktail of effector proteins into host cells that modify the activity of target host proteins. Yet, proteome-wide approaches to systematically map changes in host protein function during infection have remained challenging. Here we adapted a functional proteomics approach - Thermal-Proteome Profiling (TPP) - to systematically assess proteome-wide changes in host protein abundance and thermal stability throughout an STm infection cycle. By comparing macrophages treated with live or heat-killed STm, we observed that most host protein abundance changes occur independently of STm viability. In contrast, a large portion of host protein thermal stability changes were specific to infection with live STm. This included pronounced thermal stability changes in proteins linked to mitochondrial function (Acod1/Irg1, Cox6c, Samm50, Vdac1, and mitochondrial respiratory chain complex proteins), as well as the interferon-inducible protein with tetratricopeptide repeats, Ifit1. Integration of our TPP data with a publicly available STm-host protein-protein interaction database led us to discover that the secreted STm effector kinase, SteC, thermally destabilizes and phosphorylates the ribosomal preservation factor Serbp1. In summary, this work emphasizes the utility of measuring protein thermal stability during infection to accelerate the discovery of novel molecular interactions at the host-pathogen interface.

Authors 11

1. Marlène S. Birk Aachen Institute of Medical Microbiology

   RWTH Aachen University · Universitätsklinikum Aachen

   Affiliation as printed

   Institute of Medical Microbiology, RWTH University Hospital, Aachen 52074, Germany

2. Philipp Walch

   European Molecular Biology Laboratory · European Molecular Biology Laboratory

   Affiliation as printed

   European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany

3. Stephanie Sefried Aachen Institute of Medical Microbiology

   RWTH Aachen University · Universitätsklinikum Aachen

   Affiliation as printed

   Institute of Medical Microbiology, RWTH University Hospital, Aachen 52074, Germany

4. Jan Amelang Aachen Institute of Biochemistry and Molecular Biology

   RWTH Aachen University · Universitätsklinikum Aachen

   Affiliation as printed

   Institute of Biochemistry and Molecular Biology, RWTH University Hospital, Aachen 52074, Germany

5. Elena Buerova Aachen Institute of Biochemistry and Molecular Biology

   RWTH Aachen University · Universitätsklinikum Aachen

   Affiliation as printed

   Institute of Biochemistry and Molecular Biology, RWTH University Hospital, Aachen 52074, Germany

6. Ingrid Breuer Aachen Institute of Medical Microbiology

   RWTH Aachen University · Universitätsklinikum Aachen

   Affiliation as printed

   Institute of Medical Microbiology, RWTH University Hospital, Aachen 52074, Germany

7. Jörg Vervoorts Aachen Institute of Biochemistry and Molecular Biology

   RWTH Aachen University · Universitätsklinikum Aachen

   Affiliation as printed

   Institute of Biochemistry and Molecular Biology, RWTH University Hospital, Aachen 52074, Germany

8. Athanasios Typas

   European Molecular Biology Laboratory · European Molecular Biology Laboratory

   Affiliation as printed

   European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany

9. Mikhail M. Savitski

   European Molecular Biology Laboratory · European Molecular Biology Laboratory

   Affiliation as printed

   European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany

10. André Mateus corresponding

    Umeå University · European Molecular Biology Laboratory · European Molecular Biology Laboratory

    Affiliation as printed

    European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany; Department of Chemistry, Umeå University, Umeå 907 36, Sweden; The Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå 907 36, Sweden. Electronic address: andre.mateus@umu.se

    Department of Chemistry, Umeå University, Umeå 907 36, Sweden

    European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany

    The Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå 907 36, Sweden

11. Joel Selkrig corresponding Aachen Institute of Medical Microbiology

    RWTH Aachen University · Universitätsklinikum Aachen · European Molecular Biology Laboratory · European Molecular Biology Laboratory

    Affiliation as printed

    Institute of Medical Microbiology, RWTH University Hospital, Aachen 52074, Germany; European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany. Electronic address: jselkrig@ukaachen.de

    European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany

    Institute of Medical Microbiology, RWTH University Hospital, Aachen 52074, Germany