Oncotarget

Priority Research Papers:

Cyclin E/Cdk2-dependent phosphorylation of Mcl-1 determines its stability and cellular sensitivity to BH3 mimetics

Gaurav S. Choudhary, Trinh T. Tat, Saurav Misra, Brian T. Hill, Mitchell R. Smith, Alexandru Almasan and Suparna Mazumder _

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Oncotarget. 2015; 6:16912-16925. https://doi.org/10.18632/oncotarget.4857

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Abstract

Gaurav S. Choudhary1,6, Trinh T. Tat7, Saurav Misra2, Brian T. Hill4, Mitchell R. Smith4, Alexandru Almasan1,5 and Suparna Mazumder3,5

1 Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA

2 Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA

3 Department of Immunology, Lerner Research Institute, Taussig Cancer Institute, Cleveland Clinic, Cleveland, OH, USA

4 Department of Hematology and Oncology, Taussig Cancer Institute, Cleveland Clinic, Cleveland, OH, USA

5 Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, OH, USA

6 Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, OH, USA

7 Department of Biochemistry, Case Western Reserve University School of Medicine, Cleveland, OH, USA

Correspondence to:

Suparna Mazumder, email:

Alexandru Almasan, email:

Keywords: Mcl-1, cyclin E, Cdk2, dinaciclib ABT-737, ABT-199, CLL

Received: February 26, 2015 Accepted: July 01, 2015 Published: July 15, 2015

Abstract

Cyclin E/Cdk2 kinase activity is frequently deregulated in human cancers, resulting in impaired apoptosis. Here, we show that cyclin E/Cdk2 phosphorylates and stabilizes the pro-survival Bcl-2 family protein Mcl-1, a key cell death resistance determinant to the small molecule Bcl-2 family inhibitors ABT-199 and ABT-737, mimetics of the Bcl-2 homology domain 3 (BH3). Cyclin E levels were elevated and there was increased association of cyclin E/Cdk2 with Mcl-1 in ABT-737-resistant compared to parental cells. Cyclin E depletion in various human tumor cell-lines and cyclin E-/- mouse embryo fibroblasts showed decreased levels of Mcl-1 protein, with no change in Mcl-1 mRNA levels. In the absence of cyclin E, Mcl-1 ubiquitination was enhanced, leading to decreased protein stability. Studies with Mcl-1 phosphorylation mutants show that cyclin E/Cdk2-dependent phosphorylation of Mcl-1 residues on its PEST domain resulted in increased Mcl-1 stability (Thr92, and Thr163) and Bim binding (Ser64). Cyclin E knock-down restored ABT-737 sensitivity to acquired and inherently resistant Mcl-1-dependent tumor cells. CDK inhibition by dinaciclib resulted in Bim release from Mcl-1 in ABT-737-resistant cells. Dinaciclib in combination with ABT-737 and ABT-199 resulted in robust synergistic cell death in leukemic cells and primary chronic lymphocytic leukemia patient samples. Collectively, our findings identify a novel mechanism of cyclin E-mediated Mcl-1 regulation that provides a rationale for clinical use of Bcl-2 family and Cdk inhibitors for Mcl-1-dependent tumors.


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