title: Neddylation E2 UBE2F Promotes the Survival of Lung Cancer Cells by Activating CRL5 to Degrade NOXA via the K11 Linkage.
abstract: Purpose:Recent studies have shown that the process of protein neddylation was abnormally activated in several human cancers. However, it is unknown whether and how UBE2F, a less characterized neddylation E2, regulates lung cancer cell survival, and whether and how NOXA, a proapoptotic protein, is ubiquitylated and degraded by which E3 and via which ubiquitin linkage.Experimental Design:Methods of immunohistochemistry and immunoblotting were utilized to examine UBE2F protein expression. The biological functions of UBE2F were evaluated byin vitrocell culture andin vivoxenograft models. Thein vivocomplex formation among UBE2F-SAG-CUL5-NOXA was measured by a pulldown assay. Polyubiquitylation of NOXA was evaluated byin vivoandin vitroubiquitylation assays.Results:UBE2F is overexpressed in non-small cell lung cancer (NSCLC) and predicts poor patient survival. While UBE2F overexpression promotes lung cancer growth bothin vitroandin vivo, UBE2F knockdown selectively inhibits tumor growth. By promoting CUL5 neddylation, UBE2F/SAG/CUL5 tri-complex activates CRL5 (Cullin-RING-ligase-5) to ubiquitylate NOXA via a novel K11, but not K48, linkage for targeted proteasomal degradation. CRL5 inactivation or forced expression of K11R ubiquitin mutant caused NOXA accumulation to induce apoptosis, which is rescued by NOXA knockdown. Notably, NOXA knockdown rescues the UBE2F silencing effect, indicating a causal role of NOXA in this process. In lung cancer tissues, high levels of UBE2F and CUL5 correlate with a low level of NOXA and poor patient survival.Conclusions:By ubiquitylating and degrading NOXA through activating CRL5, UBE2F selectively promotes lung cancer cell survival and could, therefore, serve as a novel cancer target.Clin Cancer Res; 23(4); 1104-16. ©2016 AACR.
authors: Division of Radiation and Cancer Biology, Department of Radiation Oncology, University of Michigan, Ann Arbor, Michigan.
authors: Weihua
authors: W
authors: Zhou
authors: Division of Radiation and Cancer Biology, Department of Radiation Oncology, University of Michigan, Ann Arbor, Michigan.
authors: Jie
authors: J
authors: Xu
authors: Institute of Translational Medicine, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.|Children's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors: Haomin
authors: H
authors: Li
authors: Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors: Ming
authors: M
authors: Xu
authors: Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas.|Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, Texas.
authors: Zhijian J
authors: ZJ
authors: Chen
authors: Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts.
authors: Wenyi
authors: W
authors: Wei
authors: Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, New York.
authors: Zhenqiang
authors: Z
authors: Pan
authors: Division of Radiation and Cancer Biology, Department of Radiation Oncology, University of Michigan, Ann Arbor, Michigan. sunyi@umich.edu.|Institute of Translational Medicine, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.|Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University, Hangzhou, China.
authors: Yi
authors: Y
authors: Sun
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D051379:Mice; D059748:Proteolysis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics* / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics* / Q000378:metabolism; D054875:Ubiquitination / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
chemical_list: C498702:PMAIP1 protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D044763:Ubiquitin-Conjugating Enzymes; C584471:CULL-RING ligase, human; D044767:Ubiquitin-Protein Ligases; C581960:UBE2F protein, human