title: CLIPR-59 is a lipid raft-associated protein containing a cytoskeleton-associated protein glycine-rich domain (CAP-Gly) that perturbs microtubule dynamics.
abstract: We recently have identified a new cytoplasmic linker protein (CLIP), CLIPR-59, which is involved in the regulation of early endosome/trans-Golgi network dynamics. In contrast with CLIP-170, CLIPR-59 is not localized to microtubules at steady state but is associated with the trans-Golgi network and the plasma membrane. Here we show that the last 30 amino acids (C30) are sufficient for membrane targeting and that two cysteines in the C30 domain are palmitoylated. We demonstrate that CLIPR-59 is associated with lipid rafts via its C-terminal palmitoylated domain. In vitro experiments suggest that CLIPR-59 and its microtubule-binding domain alone have a better affinity for unpolymerized tubulin or small oligomers than for microtubules. In contrast with the CLIP-170 microtubule-binding domain, the CLIPR-59 microtubule-binding domain diminishes microtubule regrowth after nocodazole washout in vivo, showing that this domain can prevent microtubule polymerization. In contrast with the role of linker between membranes and microtubules that was proposed for CLIP function, CLIPR-59 thus may have an "anti-CLIP" function by preventing microtubule-raft interactions.
authors.affiliation: CNRS Unité Mixte de Recherche 144, Institut Curie Section Recherche, 75248 Paris Cedex 05, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Lallemand-Breitenbach
authors.forename: Mélanie
authors.initials: M
authors.lastname: Quesnoit
authors.forename: Virginie
authors.initials: V
authors.lastname: Braun
authors.forename: Ahmed
authors.initials: A
authors.lastname: El Marjou
authors.forename: Christian
authors.initials: C
authors.lastname: Poüs
authors.forename: Bruno
authors.initials: B
authors.lastname: Goud
authors.forename: Franck
authors.initials: F
authors.lastname: Perez
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003545:Cysteine / Q000737:chemistry; D003599:Cytoskeleton / Q000737:chemistry*; D003902:Detergents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005998:Glycine / Q000737:chemistry*; D006056:Golgi Apparatus / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D021962:Membrane Microdomains; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000502:physiology*; D008870:Microtubules / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D015739:Nocodazole / Q000494:pharmacology; D019308:Palmitic Acid / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; C449670:CLIP3 protein, human; D003902:Detergents; D008164:Luminescent Proteins; D008869:Microtubule-Associated Proteins; D049452:Green Fluorescent Proteins; D019308:Palmitic Acid; D003545:Cysteine; D015739:Nocodazole; D005998:Glycine