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A PKA survival pathway inhibited by DPT-PKI, a new specific cell permeable PKA inhibitor, is induced by T. annulata in parasitized B-lymphocytes

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Abstract

T. annulata, an intracellular pathogenic parasite of the Aplicomplexa protozoan family infects bovine B-lymphocytes and macrophages. Parasitized cells that become transformed survive and proliferate independently of exogenous growth factors. In the present study, we used the isogenic non parasitized BL3 and parasitized TBL3 B cell lines, as a model to evaluate the contribution of two-major PI3-K- and PKA-dependent anti-apoptotic pathways in the survival of T. annulata parasitized B lymphocytes. We found that T. annulata increases PKA activity, induces over-expression of the catalytic subunit and down-regulates the pro-survival phosphorylation state of Akt/PKB. Consistent with a role of PKA activation in survival, two pharmacological inhibitors H89 and KT5720 ablate PKA-dependent survival of parasitized cells. To specifically inhibit PKA pro-survival pathways we linked the DPTsh1 peptide shuttle sequence to PKI5–24 and we generated DPT-PKI, a cell permeable PKI. DPT-PKI specifically inhibited PKA activity in bovine cell extracts and, as expected, also inhibited the PKA-dependent survival of T. annulata parasitized TBL3 cells. Thus, parasite-dependent constitutive activation of PKA in TBL3 cells generates an anti-apoptotic pathway that can protect T. annulata-infected B cells from apoptosis. These results also indicate that DPT-PKI could be a powerful tool to inhibit PKA pathways in other cell types.

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Author information

Author notes

  1. Frederic Dessauge

    Present address: Unité Mixte de Recherche sur la Production du Lait-INRA-Domaine de la Motte, 35590, Saint Gilles

  2. Julien Guergnon

    Present address: Center for Cell Signaling, University of Virginia School of Medicine, 800577, West Complex MSB 7225, Charlottesville, VA, 22908, USA

  3. Gordon Langsley

    Present address: U567 INSERM/UMR 8104 CNRS/Fac de Medecine Paris V Département Maladies, Infectieuses Institut, Cochin, 22 rue Mechain, 75014, Paris, France

Authors and Affiliations

  1. Unité de Chimie Organique-Equipe Phosphatase et Laboratoire de signalisation Immunoparasitaire, URA CNRS 2581, 75015, Paris, France

    Julien Guergnon, Frederic Dessauge, Pierre Etienne Bost & Alphonse Garcia

  2. Unité de Régulation Enzymatique des Activités Cellulaires, Institut Pasteur, 25 rue du Dr Roux, 75015, Paris, France

    François Traincard

  3. Equipe Hypophyse, Physiologie de la Reproduction et des Comportements, UMR 6175 INRA-CNRS-Université de Tours-Haras Nationaux, 37380, Nouzilly, France

    Xavier Cayla

  4. Laboratoire d’Immunologie Cellulaire et Tissulaire, INSERM U543, Bâtiment CERVI Hôpital Pitié Salpétrière, 83 Bd de l’Hôpital, 75013, Paris, France

    Angelita Rebollo

  5. Laboratoire de signalisation Immunoparasitaire URA CNRS 2581, Institut Pasteur 25 rue du Dr Roux, 75015, Paris, France

    Gordon Langsley

Authors
  1. Julien Guergnon
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  2. Frederic Dessauge
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  7. Gordon Langsley
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  8. Alphonse Garcia
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Corresponding author

Correspondence to Alphonse Garcia.

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Guergnon, J., Dessauge, F., Traincard, F. et al. A PKA survival pathway inhibited by DPT-PKI, a new specific cell permeable PKA inhibitor, is induced by T. annulata in parasitized B-lymphocytes. Apoptosis 11, 1263–1273 (2006). https://doi.org/10.1007/s10495-006-7702-6

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  • DOI: https://doi.org/10.1007/s10495-006-7702-6

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