title: CBS9106-induced CRM1 degradation is mediated by cullin ring ligase activity and the neddylation pathway.
abstract: Chromosome region maintenance 1 (CRM1) mediates the nuclear export of proteins and mRNAs, and is overexpressed in various cancers. Recent studies have also reported that CRM1 protein expression is a negative prognostic factor in patients with cancer. Therefore, CRM1 is considered a potential target for anticancer therapy. Our previous study demonstrated that CBS9106, a synthetic small-molecular inhibitor of CRM1, decreases CRM1 protein through proteasomal degradation without affecting CRM1 mRNA levels. However, the mechanism by which CRM1 is degraded is not well understood. Here, we demonstrate a novel signaling pathway that plays an important role in CBS9106-induced CRM1 degradation. We found that MLN4924, a selective inhibitor of NEDD8-activating enzyme (NAE), effectively inhibits cullin neddylation and attenuates CBS9106-induced CRM1 degradation in a time- and dose-dependent manner. MLN4924 also attenuated CBS9106-induced nuclear accumulation of Ran-binding protein 1 (RanBP1), cell growth inhibition, and apoptosis. Furthermore, RNAi-mediated knockdown of neddylation pathway proteins (NEDD8 and UBA3) or cullin ring ligase (CRL) component protein (Rbx1) attenuated CRM1 protein degradation and G1 phase cell-cycle arrest by CBS9106. Knockdown of CSN5 or CAND1 also partially inhibited CBS9106-induced CRM1 degradation. These findings demonstrate that CBS9106-induced CRM1 degradation is conferred by CRL activity involving the neddylation pathway, and that this response to CBS9106 leads to cell growth inhibition and apoptosis.
authors.affiliation: CanBas Co., Ltd., Numazu, Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Saito
authors.affiliation: CanBas Co., Ltd., Numazu, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Sakakibara
authors.affiliation: CanBas Co., Ltd., Numazu, Japan.
authors.forename: Takuji
authors.initials: T
authors.lastname: Sato
authors.affiliation: CanBas Co., Ltd., Numazu, Japan.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Friedman
authors.affiliation: Dana-Farber Cancer Institute, Harvard School, Boston, Massachusetts.
authors.forename: Donald W
authors.initials: DW
authors.lastname: Kufe
authors.affiliation: Translational Genomics Research Institute (TGen), Phoenix, Arizona.
authors.forename: Daniel D
authors.initials: DD
authors.lastname: VonHoff
authors.affiliation: CanBas Co., Ltd., Numazu, Japan. takumi@canbas.co.jp.
authors.forename: Takumi
authors.initials: T
authors.lastname: Kawabe
mesh_terms: D000631:Aminopyridines / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D000075686:COP9 Signalosome Complex; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D044842:Cullin Proteins / Q000378:metabolism*; D003517:Cyclopentanes / Q000494:pharmacology; D055785:Gene Knockdown Techniques; D045325:HCT116 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D028884:Karyopherins / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism; D000075687:NEDD8 Protein; D009687:Nuclear Proteins / Q000378:metabolism; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D059748:Proteolysis / Q000187:drug effects; D011743:Pyrimidines / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D044764:Ubiquitin-Activating Enzymes / Q000235:genetics / Q000378:metabolism; D014452:Ubiquitins / Q000235:genetics / Q000378:metabolism; D000097606:Exportin 1 Protein
chemical_list: D000631:Aminopyridines; D000075686:COP9 Signalosome Complex; D002352:Carrier Proteins; D044842:Cullin Proteins; D003517:Cyclopentanes; D047908:Intracellular Signaling Peptides and Proteins; D028884:Karyopherins; D000075687:NEDD8 Protein; D009687:Nuclear Proteins; D010447:Peptide Hydrolases; D011743:Pyrimidines; D011760:Pyrrolidinones; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; D044764:Ubiquitin-Activating Enzymes; D014452:Ubiquitins; D000097606:Exportin 1 Protein; C486317:CAND1 protein, human; C568326:CBS 9106; C082860:NEDD8 protein, human; C479529:RBX1 protein, human; C091775:ran-binding protein 1; C501488:COPS5 protein, human; C491802:NAE protein, human; C539933:pevonedistat