title: Nucleophosmin (NPM1/B23) interacts with activating transcription factor 5 (ATF5) protein and promotes proteasome- and caspase-dependent ATF5 degradation in hepatocellular carcinoma cells.
abstract: Nucleophosmin (NPM1/B23) and the activating transcription factor 5 (ATF5) are both known to subject to cell type-dependent regulation. NPM1 is expressed weakly in hepatocytes and highly expressed in hepatocellular carcinomas (HCC) with a clear correlation between enhanced NPM1 expression and increased tumor grading and poor prognosis, whereas in contrast, ATF5 is expressed abundantly in hepatocytes and down-regulated in HCC. Re-expression of ATF5 in HCC inhibits cell proliferation. We report here that using an unbiased approach, tandem affinity purification (TAP) followed with mass spectrometry (MS), we identified NPM1 as a novel ATF5-interacting protein. Unlike many other NPM1-interacting proteins that interact with the N-terminal oligomerization domain of NPM1, ATF5 binds via its basic leucine zipper to the C-terminal region of NPM1 where its nucleolar localization signal is located. NPM1 association with ATF5, whose staining patterns partially overlap in the nucleoli, promotes ATF5 protein degradation through proteasome-dependent and caspase-dependent pathways. NPM1-c, a mutant NPM1 that is defective in nucleolar localization, failed to stimulate ATF5 polyubiquitination and was unable to down-regulate ATF5. NPM1 interaction with ATF5 displaces HSP70, a known ATF5-interacting protein, from ATF5 protein complexes and antagonizes its role in stabilization of ATF5 protein. NPM1-promoted ATF5 down-regulation diminished ATF5-mediated repression of cAMP-responsive element-dependent gene transcription and abrogates ATF5-induced G(2)/M cell cycle blockade and inhibition of cell proliferation in HCC cells. Our study establishes a mechanistic link between elevated NPM1 expression and depressed ATF5 in HCC and suggests that regulation of ATF5 by NPM1 plays an important role in the proliferation and survival of HCC.
authors.affiliation: Penn State College of Medicine, Hershey, Pennsylvania 17033, USA.
authors.forename: Xijun
authors.initials: X
authors.lastname: Liu
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.forename: Dongmeng
authors.initials: D
authors.lastname: Qian
authors.forename: Jenny
authors.initials: J
authors.lastname: Dai
authors.forename: Yi
authors.initials: Y
authors.lastname: An
authors.forename: Shaoyan
authors.initials: S
authors.lastname: Jiang
authors.forename: Bruce
authors.initials: B
authors.lastname: Stanley
authors.forename: Jinming
authors.initials: J
authors.lastname: Yang
authors.forename: Bin
authors.initials: B
authors.lastname: Wang
authors.forename: Xinyuan
authors.initials: X
authors.lastname: Liu
authors.forename: David X
authors.initials: DX
authors.lastname: Liu
mesh_terms: D051696:Activating Transcription Factors / Q000235:genetics / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D020169:Caspases / Q000235:genetics / Q000378:metabolism*; D059447:Cell Cycle Checkpoints / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D057809:HEK293 Cells; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D019913:Nuclear Localization Signals / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D000090243:Nucleophosmin; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D059748:Proteolysis; D020218:Response Elements / Q000235:genetics; D054875:Ubiquitination / Q000235:genetics
chemical_list: C507532:ATF5 protein, human; D051696:Activating Transcription Factors; D018840:HSP70 Heat-Shock Proteins; C000716967:NPM1 protein, human; D009363:Neoplasm Proteins; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D000090243:Nucleophosmin; D020169:Caspases; D046988:Proteasome Endopeptidase Complex