PMID- 29550478 OWN - NLM STAT- MEDLINE DCOM- 20180410 LR - 20180410 IS - 1090-2104 (Electronic) IS - 0006-291X (Linking) VI - 499 IP - 1 DP - 2018 Apr 30 TI - Cullin 3 regulates ADAMs-mediated ectodomain shedding of amphiregulin. PG - 17-23 LID - S0006-291X(18)30589-8 [pii] LID - 10.1016/j.bbrc.2018.03.097 [doi] AB - A disintegrin and metalloproteinase (ADAM) family are crucial enzymes for ectodomain shedding of multiple substrates and are involved in diverse biologic and pathologic processes. However, the molecular mechanism underlying substrate selectivity of ADAMs is poorly understood. In this study, we observed that disruption of actin polymerization by pharmacological inhibitors, latrunculin A (LatA) and cytochalasin D (CyD), induced ectodomain shedding of epidermal growth factor (EGF) family ligands. Induced shedding activity by LatA or CyD was suppressed by a metalloprotease inhibitor KB-R7785, indicating that ADAMs-mediated shedding is tightly controlled by actin cytoskeleton. We also investigated roles of cullin family, a component of cullin-RING based E3 ubiquitin ligases, in ectodomain shedding, since cullin family is implicated in the regulation of cytoskeletal dynamics. Knockdown of cullin 3 (Cul3) by a specific siRNA inhibited ectodomain shedding of amphiregulin (AREG), a member of EGF family, and responses were associated with activation of RhoA GTPase and induction of stress fiber formation. On the other hand, the RhoA inhibitor C3 transferase rescued AREG shedding reduced by Cul3 knockdown. These results describe a novel molecular mechanism of Cul3 to regulate AREG shedding by modulating cytoskeletal dynamics in a RhoA dependent manner. CI - Copyright (c) 2018. Published by Elsevier Inc. FAU - Nakayama, Hironao AU - Nakayama H AD - Division of Cell Growth and Tumor Regulation, Proteo-Science Center, Ehime University, Ehime, Japan; Department of Biochemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Toon, Shitsukawa, Ehime 791-0295, Japan; Department of Medical Science and Technology, Hiroshima International University, Higashi-hiroshima, Hiroshima 739-2695, Japan. FAU - Sakaue, Tomohisa AU - Sakaue T AD - Division of Cell Growth and Tumor Regulation, Proteo-Science Center, Ehime University, Ehime, Japan; Department of Biochemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Toon, Shitsukawa, Ehime 791-0295, Japan; Department of Cardiovascular and Thoracic Surgery, Ehime University Graduate School of Medicine, Toon, Shitsukawa, Ehime 791-0295, Japan. FAU - Maekawa, Masashi AU - Maekawa M AD - Division of Cell Growth and Tumor Regulation, Proteo-Science Center, Ehime University, Ehime, Japan; Department of Biochemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Toon, Shitsukawa, Ehime 791-0295, Japan. FAU - Fujisaki, Ayako AU - Fujisaki A AD - Division of Cell Growth and Tumor Regulation, Proteo-Science Center, Ehime University, Ehime, Japan; Department of Biochemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Toon, Shitsukawa, Ehime 791-0295, Japan. FAU - Higashiyama, Shigeki AU - Higashiyama S AD - Division of Cell Growth and Tumor Regulation, Proteo-Science Center, Ehime University, Ehime, Japan; Department of Biochemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Toon, Shitsukawa, Ehime 791-0295, Japan. Electronic address: shigeki@m.ehime-u.ac.jp. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20180321 PL - United States TA - Biochem Biophys Res Commun JT - Biochemical and biophysical research communications JID - 0372516 RN - 0 (12-O-tetradecanoylphorbol-1,3-acetate) RN - 0 (AREG protein, human) RN - 0 (Amphiregulin) RN - 0 (Bridged Bicyclo Compounds, Heterocyclic) RN - 0 (CUL3 protein, human) RN - 0 (Cullin Proteins) RN - 0 (Hydroxamic Acids) RN - 0 (Isoenzymes) RN - 0 (KB R7785) RN - 0 (RNA, Small Interfering) RN - 0 (Thiazolidines) RN - 124671-05-2 (RHOA protein, human) RN - 22144-77-0 (Cytochalasin D) RN - EC 2.4.2.- (ADP Ribose Transferases) RN - EC 2.4.2.- (exoenzyme C3, Clostridium botulinum) RN - EC 3.4.24.69 (Botulinum Toxins) RN - EC 3.4.24.86 (ADAM17 Protein) RN - EC 3.4.24.86 (ADAM17 protein, human) RN - EC 3.6.5.2 (rhoA GTP-Binding Protein) RN - NI40JAQ945 (Tetradecanoylphorbol Acetate) RN - SRQ9WWM084 (latrunculin A) RN - TE7660XO1C (Glycine) SB - IM MH - ADAM17 Protein/antagonists & inhibitors/*genetics/metabolism MH - ADP Ribose Transferases/pharmacology MH - Actin Cytoskeleton/drug effects/*metabolism/ultrastructure MH - Amphiregulin/*genetics/metabolism MH - Animals MH - Botulinum Toxins/pharmacology MH - Bridged Bicyclo Compounds, Heterocyclic/antagonists & inhibitors/pharmacology MH - Cell Line, Tumor MH - Cullin Proteins/antagonists & inhibitors/*genetics/metabolism MH - Cytochalasin D/antagonists & inhibitors/pharmacology MH - Fibroblasts/cytology/drug effects/*metabolism MH - Gene Expression Regulation MH - Glycine/analogs & derivatives/pharmacology MH - Humans MH - Hydroxamic Acids/pharmacology MH - Isoenzymes/antagonists & inhibitors/genetics/metabolism MH - RNA, Small Interfering/genetics/metabolism MH - Signal Transduction MH - Tetradecanoylphorbol Acetate/analogs & derivatives/pharmacology MH - Thiazolidines/antagonists & inhibitors/pharmacology MH - rhoA GTP-Binding Protein/antagonists & inhibitors/genetics/metabolism OTO - NOTNLM OT - Actin OT - Amphiregulin OT - Cullin 3 OT - Ectodomain shedding OT - RhoA EDAT- 2018/03/20 06:00 MHDA- 2018/04/11 06:00 CRDT- 2018/03/19 06:00 PHST- 2018/03/05 00:00 [received] PHST- 2018/03/13 00:00 [accepted] PHST- 2018/03/20 06:00 [pubmed] PHST- 2018/04/11 06:00 [medline] PHST- 2018/03/19 06:00 [entrez] AID - S0006-291X(18)30589-8 [pii] AID - 10.1016/j.bbrc.2018.03.097 [doi] PST - ppublish SO - Biochem Biophys Res Commun. 2018 Apr 30;499(1):17-23. doi: 10.1016/j.bbrc.2018.03.097. Epub 2018 Mar 21.