title: The CUL7/F-box and WD repeat domain containing 8 (CUL7/Fbxw8) ubiquitin ligase promotes degradation of hematopoietic progenitor kinase 1.
abstract: HPK1, a member of mammalian Ste20-like serine/threonine kinases, is lost in >95% pancreatic cancer through proteasome-mediated degradation. However, the mechanism of HPK1 loss has not been defined. The aims of this study are to identify the ubiquitin ligase and to examine the mechanisms that targets HPK1 degradation. We found that the CUL7/Fbxw8 ubiquitin ligase targeted HPK1 for degradation via the 26 S proteasome. The ubiquitination of HPK1 required its kinase activity and autophosphorylation. Wild-type protein phosphatase 4 (PP4), but not the phosphatase-dead PP4 mutant, PP4-RL, inhibits the interaction of Fbxw8 with HPK1 and Fbxw8-mediated ubiquitination of HPK1. In addition, we showed that Thr-355 of HPK1 is a key PP4 dephosphorylation site, through which CUL7/Fbxw8 ubiquitin ligase and PP4 regulates HPK1 stability. Knockdown of Fbxw8 restores endogenous HPK1 protein expression and inhibits cell proliferation of pancreatic cancer cells. Our study demonstrated that targeted degradation of HPK1 by the CUL7/Fbxw8 ubiquitin ligase constitutes a negative-feedback loop to restrain the activity of HPK1 and that CUL7/Fbxw8 ubiquitin ligase promotes pancreatic cancer cell proliferation. CUL7/Fbxw8 ubiquitin ligase-mediated HPK1 degradation revealed a direct link and novel role of CUL7/Fbxw8 ubiquitin ligase in the MAPK pathway, which plays a critical role in cell proliferation and differentiation.
authors.affiliation: From the Departments of Gastrointestinal Medical Oncology.
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.forename: Yue
authors.initials: Y
authors.lastname: Chen
authors.forename: Ping
authors.initials: P
authors.lastname: Lin
authors.forename: Lei
authors.initials: L
authors.lastname: Li
authors.forename: Guisheng
authors.initials: G
authors.lastname: Zhou
authors.forename: Guangchao
authors.initials: G
authors.lastname: Liu
authors.forename: Craig
authors.initials: C
authors.lastname: Logsdon
authors.forename: Jianping
authors.initials: J
authors.lastname: Jin
authors.forename: James L
authors.initials: JL
authors.lastname: Abbruzzese
authors.forename: Tse-Hua
authors.initials: TH
authors.lastname: Tan
authors.forename: Huamin
authors.initials: H
authors.lastname: Wang
mesh_terms: D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation; D044842:Cullin Proteins / Q000235:genetics / Q000378:metabolism*; D044783:F-Box Proteins / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D019169:Jurkat Cells; D020935:MAP Kinase Signaling System; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000201:enzymology* / Q000235:genetics; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D059748:Proteolysis
chemical_list: C506299:CUL7 protein, human; D044842:Cullin Proteins; D044783:F-Box Proteins; C526069:FBXW8 protein, human; D009363:Neoplasm Proteins; C102102:hematopoietic progenitor kinase 1; D017346:Protein Serine-Threonine Kinases; D010749:Phosphoprotein Phosphatases; C080406:protein phosphatase 4
keywords: CUL7 Ubiquitin Ligase; E3 Ubiquitin Ligase; Fbxw8; HPK1; MAP Kinases (MAPKs); PP4; Pancreatic Cancer; Proteasome; Protein Phosphatase; Serine Threonine Protein Kinase