title: An OBSL1-Cul7Fbxw8 ubiquitin ligase signaling mechanism regulates Golgi morphology and dendrite patterning.
abstract: The elaboration of dendrites in neurons requires secretory trafficking through the Golgi apparatus, but the mechanisms that govern Golgi function in neuronal morphogenesis in the brain have remained largely unexplored. Here, we report that the E3 ubiquitin ligase Cul7(Fbxw8) localizes to the Golgi complex in mammalian brain neurons. Inhibition of Cul7(Fbxw8) by independent approaches including Fbxw8 knockdown reveals that Cul7(Fbxw8) is selectively required for the growth and elaboration of dendrites but not axons in primary neurons and in the developing rat cerebellum in vivo. Inhibition of Cul7(Fbxw8) also dramatically impairs the morphology of the Golgi complex, leading to deficient secretory trafficking in neurons. Using an immunoprecipitation/mass spectrometry screening approach, we also uncover the cytoskeletal adaptor protein OBSL1 as a critical regulator of Cul7(Fbxw8) in Golgi morphogenesis and dendrite elaboration. OBSL1 forms a physical complex with the scaffold protein Cul7 and thereby localizes Cul7 at the Golgi apparatus. Accordingly, OBSL1 is required for the morphogenesis of the Golgi apparatus and the elaboration of dendrites. Finally, we identify the Golgi protein Grasp65 as a novel and physiologically relevant substrate of Cul7(Fbxw8) in the control of Golgi and dendrite morphogenesis in neurons. Collectively, these findings define a novel OBSL1-regulated Cul7(Fbxw8) ubiquitin signaling mechanism that orchestrates the morphogenesis of the Golgi apparatus and patterning of dendrites, with fundamental implications for our understanding of brain development.
authors.affiliation: Department of Pathology, Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Nadia
authors.initials: N
authors.lastname: Litterman
authors.forename: Yoshiho
authors.initials: Y
authors.lastname: Ikeuchi
authors.forename: Gilbert
authors.initials: G
authors.lastname: Gallardo
authors.forename: Brenda C
authors.initials: BC
authors.lastname: O'Connell
authors.forename: Mathew E
authors.initials: ME
authors.lastname: Sowa
authors.forename: Steven P
authors.initials: SP
authors.lastname: Gygi
authors.forename: J Wade
authors.initials: JW
authors.lastname: Harper
authors.forename: Azad
authors.initials: A
authors.lastname: Bonni
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D044842:Cullin Proteins / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D003712:Dendrites / Q000201:enzymology* / Q000502:physiology; D018274:Electroporation; D044783:F-Box Proteins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000201:enzymology* / Q000502:physiology; D000077614:Golgi Matrix Proteins; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D009024:Morphogenesis; D040901:Proteomics; D051381:Rats; D015398:Signal Transduction; D014162:Transfection; D054875:Ubiquitination
chemical_list: D044842:Cullin Proteins; D003598:Cytoskeletal Proteins; D044783:F-Box Proteins; D000077614:Golgi Matrix Proteins; C491577:Gorasp1 protein, rat; D008565:Membrane Proteins; C522001:OBSL1 protein, rat