Drosophila Nedd4, a ubiquitin ligase, is recruited by Commissureless to control cell surface levels of the roundabout receptor.
Crossing the midline produces changes in axons such that they are no longer attracted to the midline. In Drosophila, Roundabout reaches high levels on axons once they have crossed the midline, and this prohibits recrossing. Roundabout protein levels are regulated by Commissureless. We show that Commissureless binds to and is regulated by the ubiquitin ligase DNedd4. We further show that the ability of Commissureless to regulate Roundabout protein levels requires an intact DNedd4 binding site and ubiquitin acceptor sites within the Commissureless protein. The ability of Commissureless to regulate Robo in the embryo also requires a Commissureless/DNedd4 interaction. Our results show that changes in axonal sensitivity to external cues during pathfinding across the midline makes use of ubiquitin-dependent mechanisms to regulate transmembrane protein levels.
Myat
Anna
A
Molecular Neurobiology Group, MRC Centre for Developmental Neurobiology, King's College London, New Hunt's House, Guy's Campus, London SE1 1UL, United Kingdom.
Henry
Pauline
P
McCabe
Veronica
V
Flintoft
Louisa
L
Rotin
Daniela
D
Tear
Guy
G
D000818:Animals; D001369:Axons / Q000201:enzymology* / Q000648:ultrastructure; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000302:isolation & purification*; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000166:cytology / Q000196:embryology / Q000201:enzymology*; D056827:Endosomal Sorting Complexes Required for Transport; D007839:Functional Laterality / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D020439:Growth Cones / Q000378:metabolism / Q000648:ultrastructure; D007424:Intracellular Fluid / Q000378:metabolism; D008025:Ligases / Q000235:genetics / Q000302:isolation & purification*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D000075702:Nedd4 Ubiquitin Protein Ligases; D009419:Nerve Tissue Proteins; D009420:Nervous System / Q000166:cytology / Q000196:embryology / Q000201:enzymology*; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D011956:Receptors, Cell Surface / Q000378:metabolism; D011971:Receptors, Immunologic / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D022161:Transport Vesicles / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases; D000097905:Roundabout Proteins
D002135:Calcium-Binding Proteins; D018076:DNA, Complementary; D029721:Drosophila Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D008025:Ligases; D008565:Membrane Proteins; D000075702:Nedd4 Ubiquitin Protein Ligases; D009419:Nerve Tissue Proteins; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D025801:Ubiquitin; D044767:Ubiquitin-Protein Ligases; D000097905:Roundabout Proteins; C110665:comm protein, Drosophila