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The apoptotic v-cyclin–CDK6 complex phosphorylates and inactivates Bcl-2

Nature Cell Biology volume 2, pages 819–825 (2000)Cite this article

Abstract

v-cyclin encoded by Kaposi's sarcoma herpesvirus/human herpesvirus 8 (KSHV or HHV8) associates with cellular cyclin-dependent kinase 6 (CDK6) to form a kinase complex that promotes cell-cycle progression, but can also induce apoptosis in cells with high levels of CDK6. Here we show that whereas HHV8-encoded v-Bcl-2 protects against this apoptosis, cellular Bcl-2 has lost its anti-apoptotic potential as a result of an inactivating phosphorylation in its unstructured loop region. Moreover, we identify Bcl-2 as a new substrate for v-cyclin–CDK6 in vitro, and show that it is present in a complex with CDK6 in cell lysates. A Bcl-2 mutant with a S70A S87A double substitution in the loop region is not phosphorylated and provides resistance to apoptosis, indicating that inactivation of Bcl-2 by v-cyclin–CDK6 may be required for the observed apoptosis. Furthermore, the identification of phosphorylated Bcl-2 in HHV8-positive Kaposi's sarcoma indicates that HHV8-mediated interference with host apoptotic signalling pathways may encourage the development of Kaposi's sarcoma.

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Figure 1: The inability of Bcl-2 to protect from cell death is specific to v-cyclin–CDK6-induced apoptosis.
Figure 2: Bcl-2 is phosphorylated in cells co-expressing v-cyclin and CDK6.
Figure 3: Bcl-2 phosphorylation in cells expressing v-cyclin and CDK6 is not mediated by a G2/M-activated JNK1 pathway.
Figure 4: Complex formation and direct phosphorylation of Bcl-2 by CDK6 in conjunction with v-cyclin.
Figure 5: Bcl-2 is phosphorylated by v-cyclin–CDK6 at Ser70 and Ser87 and thereby inactivated.
Figure 6: Phosphorylation of Bcl-2 in HHV8-positive BC-3 PEL cells and in a primary Kaposi's sarcoma.

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Acknowledgements

We thank N. Kalkkinen and J. Helin for the MALDI-TOF analysis, and E. E. Kaaya and C. Massambu (Muhimbili Univ. College, Dar-es-Salam, Tanzania) for the Kaposi's sarcoma biopsy material. We are also grateful to E. Cesarman, M. Hardwick, C. Sherr, S. Mittnacht, B. Chandran, L. Andersson, P. Moore, Y. Chang, S. Leppä and M. Eriksson for reagents, M. Schoultz for help with fluorescence-activated cell-sorter analysis, and members of the Mäkelä laboratory for discussions and suggestions. B. Tjäder provided excellent technical assistance. This study was supported by grants from the Academy of Finland, Univ. Helsinki, Helsinki Univ. Central Hospital EVO funds, Finnish Cancer Society, Finnish Cancer Institute, Sigrid Juselius Foundation, and the BIOMED-2 Concerted Action (CA) on the `Pathogenesis of AIDS Kaposi's sarcoma' (contract no. BMH4-97-2302).

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  1. Kazuhito Yamamoto

    Present address: The First Department of Internal Medicine, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi, 466-8550, Japan

Authors and Affiliations

  1. Haartman Institute & Biocentrum Helsinki, University of Helsinki, PO Box 21, Haartmaninkatu 3, Helsinki, 00014, Finland

    Päivi M. Ojala & Tomi P. Mäkelä

  2. HUCH Laboratory Diagnostics, Helsinki University Central Hospital, PO Box 401, HYKS, 00029, Finland

    Päivi M. Ojala & Tomi P. Mäkelä

  3. Departments of Pathology and Medicine, Harvard Medical School and Dana-Farber Cancer Institute, 1 Jimmy Fund Way, Boston, 02115, Massachusetts, USA

    Kazuhito Yamamoto & Stanley J. Korsmeyer

  4. Department of Pathology and Oncology, Immunopathology Laboratory, Karolinska Institute/Hospital, Stockholm, S-171 76, Sweden

    Esmeralda Castaños-Vélez & Peter Biberfeld

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Ojala, P., Yamamoto, K., Castaños-Vélez, E. et al. The apoptotic v-cyclin–CDK6 complex phosphorylates and inactivates Bcl-2. Nat Cell Biol 2, 819–825 (2000). https://doi.org/10.1038/35041064

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