title: Nuclear FAM21 participates in NF-κB-dependent gene regulation in pancreatic cancer cells.
abstract: The pentameric WASH complex is best known for its role in regulating receptor trafficking from retromer-rich endosomal subdomains. FAM21 functions to stabilize the WASH complex through its N-terminal head domain and localizes it to endosomes by directly binding the retromer through its extended C-terminal tail. Herein, we used affinity purification combined with mass spectrometry to identify additional FAM21-interacting proteins. Surprisingly, multiple components of the nuclear factor κB (NF-κB) pathway were identified, including the p50 and p65 (RelA) NF-κB subunits. We show that FAM21 interacts with these components and regulates NF-κB-dependent gene transcription at the level of p65 chromatin binding. We further demonstrate that FAM21 contains a functional monopartite nuclear localization signal sequence (NLS) as well as a CRM1/exportin1-dependent nuclear export signal (NES), both of which work jointly with the N-terminal head domain and C-terminal retromer recruitment domain to regulate FAM21 cytosolic and nuclear subcellular localization. Finally, our findings indicate that FAM21 depletion sensitizes pancreatic cancer cells to gemcitabine and 5-fluorouracil. Thus, FAM21 not only functions as an integral component of the cytoplasmic WASH complex, but also modulates NF-κB gene transcription in the nucleus.
authors.affiliation: Division of Oncology Research and Schulze Center for Novel Therapeutics, Mayo Clinic College of Medicine, Rochester, MN 55905, USA Department of Pathophysiology, Qiqihar Medical University, Qiqihar, Heilongjiang 161006, China.
authors.forename: Zhi-Hui
authors.initials: ZH
authors.lastname: Deng
authors.affiliation: Division of Oncology Research and Schulze Center for Novel Therapeutics, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Gomez
authors.affiliation: Division of Oncology Research and Schulze Center for Novel Therapeutics, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
authors.forename: Douglas G
authors.initials: DG
authors.lastname: Osborne
authors.affiliation: Division of Oncology Research and Schulze Center for Novel Therapeutics, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
authors.forename: Christine A
authors.initials: CA
authors.lastname: Phillips-Krawczak
authors.affiliation: Division of Oncology Research and Schulze Center for Novel Therapeutics, Mayo Clinic College of Medicine, Rochester, MN 55905, USA Key Laboratory of Biotechnology and Pharmaceutical Engineering, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China zhang.jinsan@mayo.edu billadeau.daniel@mayo.edu.
authors.forename: Jin-San
authors.initials: JS
authors.lastname: Zhang
authors.affiliation: Division of Oncology Research and Schulze Center for Novel Therapeutics, Mayo Clinic College of Medicine, Rochester, MN 55905, USA Department of Immunology, College of Medicine, Mayo Clinic, Rochester, MN 55905, USA zhang.jinsan@mayo.edu billadeau.daniel@mayo.edu.
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Billadeau
mesh_terms: D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000235:genetics; D003593:Cytoplasm / Q000378:metabolism; D003841:Deoxycytidine / Q000008:administration & dosage / Q000031:analogs & derivatives; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D019913:Nuclear Localization Signals / Q000235:genetics; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D028044:Phosphate-Binding Proteins; D011485:Protein Binding / Q000235:genetics; D011506:Proteins / Q000235:genetics*; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism*; D000093542:Gemcitabine
chemical_list: D002843:Chromatin; D008840:Microfilament Proteins; D016328:NF-kappa B; D019913:Nuclear Localization Signals; D028044:Phosphate-Binding Proteins; D011506:Proteins; C498143:RELA protein, human; D051996:Transcription Factor RelA; C549919:WASH protein, human; C000605267:WASHC2C protein, human; D003841:Deoxycytidine; D000093542:Gemcitabine
keywords: FAM21; NF-κB; Pancreatic cancer; Retromer; WASH complex