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No evidence for immunoproteasomes in chicken lymphoid organs and activated lymphocytes

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Abstract

The proteasome is the main protein-degrading machine within the cell, producing ligands for MHC class I molecules. It is a cylindrical multicatalytic protease complex, and the catalytic activity is mediated by the three subunits β1, β2, and β5 which possess caspase-, trypsin-, and chymotrypsin-like activities, respectively. By stimulation with interferon (IFN)-γ the replacement of these subunits by β1i, β2i, and β5i is induced leading to formation of immunoproteasomes with altered proteolytic and antigen processing properties. The genes coding for these immunosubunits are restricted to jawed vertebrates but have so far not been found in the genomes of birds, e.g., chicken, turkey, quail, black grouse and zebra finch. However, the chicken genome sequences are not completely assigned; therefore, we investigated the presence of immunoproteasome on protein level. 20S proteasome was purified from the chicken brain, blood, spleen, and bursa of Fabricius, followed by separation via two-dimensional (2D) gel electrophoresis. We analyzed the protein spots derived from the spleen and brain by mass spectrometry and could identify all 14 proteasomal subunits, but there were no differences detectable in the spot patterns. Moreover, we stimulated the chicken spleen cells with phorbol 12-myristate 13-acetate (PMA) and ionomycin aiming at the induction of immunoproteasome, but in spite of the induction of proliferation and IFN-γ, no evidence for immunoproteasome formation in chicken could be obtained. This result was substantiated by the finding that 20S proteasomes isolated from immune and non-immune tissues showed very similar peptidolytic activities. Taken together, our results indicate that chicken lack immunoproteasomes also on protein level.

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Acknowledgments

We thank Gerardo Alvarez Salinas, Gerhard Gröttrup, and Lars Krieger for their help with proteasome purification and Thomas Göbel (LMU Munich) and Jim Kaufman (Cambridge University) for helpful advice concerning the spleen stimulation experiments. We are grateful to Julia Koerner and Michael Basler for help with flow cytometric analyses. The team of the animal research facility of Konstanz University is acknowledged for assistance during animal experimentation. This work was supported by a grant from the German Research Foundation (Nr. GR 1517/14-1) and a grant from the Swiss National Science Foundation (Nr. 31003A_138451).

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  1. Department of Immunology, University of Konstanz, Universitaetsstrasse 10, 78464, Konstanz, Germany

    Sonja Erath & Marcus Groettrup

  2. Biotechnology Institute Thurgau at the University of Konstanz, Unterseestrasse 47, 8280, Kreuzlingen, Switzerland

    Marcus Groettrup

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  1. Sonja Erath
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Correspondence to Marcus Groettrup.

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Erath, S., Groettrup, M. No evidence for immunoproteasomes in chicken lymphoid organs and activated lymphocytes. Immunogenetics 67, 51–60 (2015). https://doi.org/10.1007/s00251-014-0814-1

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  • DOI: https://doi.org/10.1007/s00251-014-0814-1

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