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Mutant B-RAF signaling and cyclin D1 regulate Cks1/S-phase kinase-associated protein 2-mediated degradation of p27Kip1 in human melanoma cells

Oncogene volume 26, pages 1056–1066 (2007)Cite this article

Abstract

Levels of cyclins and cyclin-dependent kinase (Cdk) inhibitors are tightly controlled during normal cell proliferation and are frequently dysregulated in cancerous cells. In melanoma, cyclin D1 is highly expressed and downregulation of the Cdk inhibitor, p27Kip1, is associated with a poor prognosis. Mutant B-RAF is frequently expressed in melanoma and overrides growth factor and matrix adhesion control of cyclin D1 and p27Kip1 levels in human melanocytes. Here, we demonstrate that p27Kip1 expression is regulated by multiple mechanisms in melanoma cells. B-RAF regulates p27Kip1 mRNA abundance independently of cyclin D1. Additionally, B-RAF and cyclin D1 control the levels of S-phase kinase-associated protein 2 (Skp2) that directs ubiquitin-mediated proteolysis of p27Kip1. The cofactor for Skp2, Cdc kinase subunit 1 (Cks1) controls levels of Skp2 in melanoma cells and acts jointly with Skp2 to regulate p27Kip1 levels. Importantly, expression of Cks1 is regulated by B-RAF and cyclin D1 at the mRNA level. Reduced Cks1 or Skp2 expression and enhanced p27Kip1 levels inhibit melanoma cell growth. In summary, p27Kip1 expression in melanoma is regulated by B-RAF at the mRNA level, and via B-RAF and cyclin D1 control of Cks1/Skp2-mediated proteolysis.

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Abbreviations

Cdk:

cyclin-dependent kinases

Cks1:

cdc kinase subunit 1

NHEM:

normal human epidermal melanocytes

qRT–PCR:

real-time quantitative reverse transcriptase–polymerase chain reaction

Rb:

retinoblastoma

RNAi:

RNA interference

SCF:

Skp1/Cul1/F-box

Skp2:

S-phase kinase-associated protein 2

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Acknowledgements

The help of the following researchers/facilities is gratefully acknowledged: The Birth Place at Albany Medical Center provided research samples; Dr Meenhard Herlyn for cell lines; Dr Mark Bond donated the Skp2 adenoviruses; Dr Kevin Pumiglia and Patrick Bryant assisted with the adenoviral amplification and purification; Drs Nakayama and Matsumoto provided the p27Kip1(T187A) cDNA; and Dr Thomas Friedrich provided helpful comments on the work. The Aplin lab is supported by National Institutes of Health Grant GM067893. The authors have no competing financial interests.

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  1. Center for Cell Biology and Cancer Research, 47 New Scotland Avenue, Albany Medical College, Albany, NY, USA

    K V Bhatt, R Hu, L S Spofford & A E Aplin

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Bhatt, K., Hu, R., Spofford, L. et al. Mutant B-RAF signaling and cyclin D1 regulate Cks1/S-phase kinase-associated protein 2-mediated degradation of p27Kip1 in human melanoma cells. Oncogene 26, 1056–1066 (2007). https://doi.org/10.1038/sj.onc.1209861

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