title: Rb and FZR1/Cdh1 determine CDK4/6-cyclin D requirement in C. elegans and human cancer cells.
abstract: Cyclin-dependent kinases 4 and 6 (CDK4/6) in complex with D-type cyclins promote cell cycle entry. Most human cancers contain overactive CDK4/6-cyclin D, and CDK4/6-specific inhibitors are promising anti-cancer therapeutics. Here, we investigate the critical functions of CDK4/6-cyclin D kinases, starting from an unbiased screen in the nematode Caenorhabditis elegans. We found that simultaneous mutation of lin-35, a retinoblastoma (Rb)-related gene, and fzr-1, an orthologue to the APC/C co-activator Cdh1, completely eliminates the essential requirement of CDK4/6-cyclin D (CDK-4/CYD-1) in C. elegans. CDK-4/CYD-1 phosphorylates specific residues in the LIN-35 Rb spacer domain and FZR-1 amino terminus, resembling inactivating phosphorylations of the human proteins. In human breast cancer cells, simultaneous knockdown of Rb and FZR1 synergistically bypasses cell division arrest induced by the CDK4/6-specific inhibitor PD-0332991. Our data identify FZR1 as a candidate CDK4/6-cyclin D substrate and point to an APC/C(FZR1) activity as an important determinant in response to CDK4/6-inhibitors.
authors.affiliation: Developmental Biology, Faculty of Sciences, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Inge
authors.initials: I
authors.lastname: The
authors.affiliation: Developmental Biology, Faculty of Sciences, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Suzan
authors.initials: S
authors.lastname: Ruijtenberg
authors.affiliation: Cell Biology, Faculty of Sciences, Department of Biology, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.
authors.forename: Benjamin P
authors.initials: BP
authors.lastname: Bouchet
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics Group, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Alba
authors.initials: A
authors.lastname: Cristobal
authors.affiliation: Developmental Biology, Faculty of Sciences, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Martine B W
authors.initials: MB
authors.lastname: Prinsen
authors.affiliation: Developmental Biology, Faculty of Sciences, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Tim
authors.initials: T
authors.lastname: van Mourik
authors.affiliation: Hematologic Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, Massachusetts 02215, USA.
authors.forename: John
authors.initials: J
authors.lastname: Koreth
authors.affiliation: Department of Pathology and Laboratory Medicine, Boston University School of Medicine and Boston Medical Center, 670 Albany Street, Boston, MA, USA.
authors.forename: Huihong
authors.initials: H
authors.lastname: Xu
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics Group, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Albert J R
authors.initials: AJ
authors.lastname: Heck
authors.affiliation: Cell Biology, Faculty of Sciences, Department of Biology, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.
authors.forename: Anna
authors.initials: A
authors.lastname: Akhmanova
authors.affiliation: Hubrecht Institute, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands.
authors.forename: Edwin
authors.initials: E
authors.lastname: Cuppen
authors.affiliation: Developmental Biology, Faculty of Sciences, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Mike
authors.initials: M
authors.lastname: Boxem
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics Group, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Javier
authors.initials: J
authors.lastname: Muñoz
authors.affiliation: Developmental Biology, Faculty of Sciences, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Sander
authors.initials: S
authors.lastname: van den Heuvel
mesh_terms: D000818:Animals; D001483:Base Sequence; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000235:genetics; D064200:Cdh1 Proteins / Q000235:genetics / Q000378:metabolism*; D002453:Cell Cycle / Q000502:physiology*; D045744:Cell Line, Tumor; D056741:Cyclin D / Q000378:metabolism; D051358:Cyclin-Dependent Kinase 4 / Q000378:metabolism; D051361:Cyclin-Dependent Kinase 6 / Q000378:metabolism; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics; D016160:Retinoblastoma Protein / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA
chemical_list: D029742:Caenorhabditis elegans Proteins; D064200:Cdh1 Proteins; D056741:Cyclin D; C577584:FZR1 protein, human; D046912:Multiprotein Complexes; D012097:Repressor Proteins; D016160:Retinoblastoma Protein; C116485:lin-35 protein, C elegans; D051358:Cyclin-Dependent Kinase 4; D051361:Cyclin-Dependent Kinase 6