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hSMG-1 is a granzyme B-associated stress-responsive protein kinase

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Abstract

Granzyme B plays a key role in cell-mediated programmed cell death. We previously demonstrated that p53 is a functional determinant in the granzyme B-induced cytotoxic T-lymphocyte response. However, the pathways leading to activation of p53 by granzyme B remain incompletely understood. We now demonstrate that granzyme B-induced DNA damage signaling as revealed by histone H2AX phosphorylation and subsequent activation of the stress kinase CHK2. Confocal microscopy analysis indicates that granzyme B treatment of tumor cells induced an early translocation of endonuclease caspase-activated DNase. DNA microarray-based global transcriptional profiling and RT-PCR indeed revealed genes related to DNA damage. Among these genes, hSMG-1, a genotoxic stress-activated protein, was constantly upregulated in tumor cells following granzyme B treatment. Knockdown of the hSMG-1 gene in T1 tumor target cell line resulted in a significant inhibition of granzyme B- and CTL-induced killing. Our data suggest that granzyme B may exert cell death through DNA damage signaling and uncover a novel molecular link between the DNA damage pathway and granzyme B-induced cell death.

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Abbreviations

ATM:

Ataxia-telangiectasia mutated

CHK2:

Checkpoint kinase 2

CTL:

Cytotoxic T lymphocyte

DSBs:

Double-strand breaks

GrB:

Granzyme B

H2AX:

Histone 2AX

ICAD:

Inhibitor of CAD

PIKK:

Phosphatidylinositol 3-kinase-related kinases

SLO:

Streptolysin-O

hSMG-1:

Human suppressor of morphogenesis in genitalia-1

TNF:

Tumor necrosis factor

TRAIL:

TNF-related apoptosis

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Acknowledgements

We thank Florence Faure for T1 cells and Lisa Bain for editing the manuscript. This work was supported by grants from the INSERM, the “Association pour la Recherche contre le Cancer” (ARC, grant 3922) and the “Ligue contre le Cancer” (SR2005-430, comité des Hauts de Seine), and the “Agence Nationale de la Recherche”.

Data availability

The microarray data related to this paper have been submitted to the Array Express data repository at the European Bioinformatics Institute (http://www.ebi.ac.uk/arrayexpress/) under the accession number XXX.

Authorships

Meslin F., Hamaï A., Rosselli F., Wemhoff G., Thiery J., and Chouaib S. designed and performed the research project, analyzed data, and wrote the manuscript; Richon C. performed technical assistance for the DNA microarray assay; Jalil A. performed confocal microscopy analysis; Mlecnik B. and Galon J. contributed to the DNA microarray analysis.

Disclosures

The authors declare no financial conflict of interests.

Author information

Authors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale (INSERM) U753; Laboratoire d’immunologie des tumeurs humaines: interaction effecteurs cytotoxiques-système tumoral, Institut Gustave Roussy PR1 and IFR 54, 94805, Villejuif Cedex, France

    Franck Meslin, Ahmed Hamaï, Abdelali Jalil, Jerome Thiery & Salem Chouaib

  2. Institut National de la Santé et de la Recherche Médicale (INSERM) U872, Centre de Recherche des Cordeliers, 75006, Paris, France

    Bernhard Mlecnik & Jerome Galon

  3. Centre National de la Recherche Scientifique (CNRS) FRE2939, Laboratoire de stabilité génétique et oncogenèse, Institut Gustave Roussy PR2 and IFR 54, 94805, Villejuif Cedex, France

    Filippo Rosselli

  4. Functional Genomic Unit, Institut Gustave Roussy PR2 and IFR 54, 94805, Villejuif Cedex, France

    Catherine Richon

  5. Functional Genomics, Sigma–Aldrich, 2909 Laclede Avenue, St. Louis, MO, 63103, USA

    Gregory Wemhoff

  6. Immune Disease Institute, Harvard Medical School, 200 Longwood Avenue, Boston, MA, 02115, USA

    Jerome Thiery

Authors
  1. Franck Meslin
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  2. Ahmed Hamaï
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  3. Bernhard Mlecnik
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  4. Filippo Rosselli
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  5. Catherine Richon
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  6. Abdelali Jalil
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  7. Gregory Wemhoff
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  8. Jerome Thiery
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  9. Jerome Galon
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  10. Salem Chouaib
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Corresponding author

Correspondence to Salem Chouaib.

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Meslin, F., Hamaï, A., Mlecnik, B. et al. hSMG-1 is a granzyme B-associated stress-responsive protein kinase. J Mol Med 89, 411–421 (2011). https://doi.org/10.1007/s00109-010-0708-0

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  • DOI: https://doi.org/10.1007/s00109-010-0708-0

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