title: Small molecule targeting Cdc42-intersectin interaction disrupts Golgi organization and suppresses cell motility.
abstract: Signaling through the Rho family of small GTPases has been intensely investigated for its crucial roles in a wide variety of human diseases. Although RhoA and Rac1 signaling pathways are frequently exploited with the aid of effective small molecule modulators, studies of the Cdc42 subclass have lagged because of a lack of such means. We have applied high-throughput in silico screening and identified compounds that are able to fit into the surface groove of Cdc42, which is critical for guanine nucleotide exchange factor binding. Based on the interaction between Cdc42 and intersectin (ITSN), a specific Cdc42 guanine nucleotide exchange factor, we discovered compounds that rendered ITSN-like interactions in the binding pocket. By using in vitro binding and imaging as well as biochemical and cell-based assays, we demonstrated that ZCL278 has emerged as a selective Cdc42 small molecule modulator that directly binds to Cdc42 and inhibits its functions. In Swiss 3T3 fibroblast cultures, ZCL278 abolished microspike formation and disrupted GM130-docked Golgi structures, two of the most prominent Cdc42-mediated subcellular events. ZCL278 reduces the perinuclear accumulation of active Cdc42 in contrast to NSC23766, a selective Rac inhibitor. ZCL278 suppresses Cdc42-mediated neuronal branching and growth cone dynamics as well as actin-based motility and migration in a metastatic prostate cancer cell line (i.e., PC-3) without disrupting cell viability. Thus, ZCL278 is a small molecule that specifically targets Cdc42-ITSN interaction and inhibits Cdc42-mediated cellular processes, thus providing a powerful tool for research of Cdc42 subclass of Rho GTPases in human pathogenesis, such as those of cancer and neurological disorders.
authors.affiliation: Department of Anatomy and Cell Biology, Brody School of Medicine, East Carolina University, Greenville, NC 27834, USA.
authors.forename: Amy
authors.initials: A
authors.lastname: Friesland
authors.forename: Yaxue
authors.initials: Y
authors.lastname: Zhao
authors.forename: Yan-Hua
authors.initials: YH
authors.lastname: Chen
authors.forename: Lie
authors.initials: L
authors.lastname: Wang
authors.forename: Huchen
authors.initials: H
authors.lastname: Zhou
authors.forename: Qun
authors.initials: Q
authors.lastname: Lu
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D006056:Golgi Apparatus / Q000187:drug effects / Q000502:physiology; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009474:Neurons / Q000187:drug effects / Q000648:ultrastructure; D054730:Protein Interaction Domains and Motifs / Q000187:drug effects; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction / Q000187:drug effects; D041701:Swiss 3T3 Cells; D014584:User-Computer Interface; D014945:Wound Healing / Q000187:drug effects; D020764:cdc42 GTP-Binding Protein / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000235:genetics / Q000502:physiology
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; C115919:intersectin 1; D020764:cdc42 GTP-Binding Protein