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Uncovering residues that regulate cyclin D1 proteasomal degradation

Oncogene volume 26, pages 5098–5106 (2007)Cite this article

Abstract

Cyclin D1 regulates G1 cell-cycle progression and is aberrantly expressed in carcinogenesis. Proteasomal degradation of cyclin D1 was highlighted as a cancer chemopreventive mechanism. To understand this mechanism better, residues responsible for degradation and ubiquitination of cyclin D1 were investigated. Eighteen lysines in cyclin D1 had single, double or multiple mutations engineered before transfection into BEAS-2B human bronchial epithelial (HBE) cells to evaluate stabilities after all-trans-retinoic acid (RA) or cycloheximide treatments. Specific mutations stabilized cyclin D1, including substitutions of lysines surrounding the cyclin box domain that inhibited RA-mediated degradation and extended the cyclin D1 half-life. Mutation of all cyclin D1 lysines blocked polyubiquitination. N-terminus (but not C-terminus) modification stabilized cyclin D1. Ubiquitination-resistant mutants preferentially localized cyclin D1 to the nucleus, directly implicating subcellular localization in regulating cyclin D1 degradation. Taken together, these findings uncover specific residues conferring ubiquitination of cyclin D1. These provide a mechanistic basis for proteasomal degradation of cyclin D1.

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Acknowledgements

This work was supported by National Institutes of Health and National Cancer Institute grants RO1-CA087546 (ED) and RO1-CA111422 (ED) and by the Samuel Waxman Cancer Research Foundation (ED). We thank Ms Ann M Lavanway (Dartmouth College) for assistance with confocal microscopy.

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, NH, USA

    Q Feng, D Sekula, S J Freemantle & E Dmitrovsky

  2. Institut für Molekularbiologie und Tumorforschung (IMT), Philipps-Universität Marburg, Emil-Mannkopff-Strasse 2, Marburg, Germany

    R Müller

  3. Department of Medicine, Dartmouth Medical School, Hanover, NH, USA

    E Dmitrovsky

  4. Norris Cotton Cancer Center, Dartmouth Medical School, Hanover, NH, USA

    E Dmitrovsky

  5. Dartmouth-Hitchcock Medical Center, Lebanon, NH, USA

    E Dmitrovsky

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  1. Q Feng
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Correspondence to E Dmitrovsky.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Feng, Q., Sekula, D., Müller, R. et al. Uncovering residues that regulate cyclin D1 proteasomal degradation. Oncogene 26, 5098–5106 (2007). https://doi.org/10.1038/sj.onc.1210309

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  • DOI: https://doi.org/10.1038/sj.onc.1210309

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