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Related Experiment Videos

KSHV vFLIP binds to IKK-gamma to activate IKK.

Nigel Field1, Walter Low, Mark Daniels

  • 1Department of Immunology and Molecular Pathology, University College London, Windeyer Institute, 46 Cleveland St, London W1T 2AH, UK.

Journal of Cell Science
|August 2, 2003
PubMed
Summary

Kaposi's sarcoma-associated herpesvirus (KSHV) viral FLIP protein (vFLIP) interacts with IKK components, activating NF-kappaB. This interaction, involving Hsp90, promotes survival in KSHV-infected lymphoma cells.

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Area of Science:

  • Oncology
  • Virology
  • Molecular Biology

Background:

  • Kaposi's sarcoma-associated herpesvirus (KSHV) viral FLIP protein (vFLIP) modulates host cell pathways.
  • vFLIP is known to inhibit apoptosis and activate NF-kappaB signaling via IkappaB kinase (IKK).

Purpose of the Study:

  • To elucidate the molecular interactions of vFLIP within KSHV-infected cells.
  • To investigate the role of vFLIP-IKK complex in primary effusion lymphoma (PEL) cell survival.

Main Methods:

  • Yeast two-hybrid screening to identify vFLIP interacting partners.
  • Mammalian and bacterial expression of IKKgamma fragments to map vFLIP binding region.
  • Immunoprecipitation and mass spectrometry in PEL cells to identify vFLIP-associated proteins.
  • Gel filtration chromatography to analyze vFLIP-IKK complex formation.

Related Experiment Videos

  • Inhibition studies using geldanamycin, an Hsp90 inhibitor.

    • Main Results:

    • IKKgamma was identified as a vFLIP interacting partner, with binding localized to the central CCR3/4 domain of IKKgamma.
    • vFLIP immunoprecipitates from PEL cells revealed associated IKK subunits (IKKalpha, beta, gamma) and Hsp90.
    • vFLIP co-eluted and co-precipitated with an activated IKK complex in the cytoplasm of PEL cells.
    • Hsp90 inhibition by geldanamycin reduced vFLIP-induced IKK kinase activity and induced PEL cell death.

    Conclusions:

    • vFLIP forms a complex with IKK subunits and Hsp90 in KSHV-infected PEL cells.
    • vFLIP-mediated activation of IKK contributes to the survival of PEL cells.
    • Targeting the vFLIP-IKK-Hsp90 axis may represent a therapeutic strategy for KSHV-associated malignancies.