Doublecortin interacts with mu subunits of clathrin adaptor complexes in the developing nervous system.
Doublecortin is a microtubule-associated protein required for normal corticogenesis in the developing brain. We carried out a yeast two-hybrid screen to identify interacting proteins. One of the isolated clones encodes the mu1 subunit of the adaptor complex AP-1 involved in clathrin-dependent protein sorting. We found that Doublecortin also interacts in yeast with mu2 from the AP-2 complex. Mutagenesis and pull-down experiments showed that these interactions were mediated through a tyrosine-based sorting signal (YLPL) in the C-terminal part of Doublecortin. The functional relevance of these interactions was suggested by the coimmunoprecipitation of Doublecortin with AP-1 and AP-2 from mouse brain extracts. This interaction was further supported by RNA in situ hybridization and immunofluorescence studies. Taken together these data indicate that a certain proportion of Doublecortin interacts with AP-1 and/or AP-2 in vivo and are consistent with a potential involvement of Doublecortin in protein sorting or vesicular trafficking.
Friocourt
G
G
Laboratoire de Génétique et Physiopathologie des retards mentaux, ICGM, INSERM, CHU, Cochin, 24, rue du Faubourg Saint Jacques, Paris, 75014, France.
Chafey
P
P
Billuart
P
P
Koulakoff
A
A
Vinet
M C
MC
Schaar
B T
BT
McConnell
S K
SK
Francis
F
F
Chelly
J
J
D033961:Adaptor Protein Complex 1; D033962:Adaptor Protein Complex 2; D033963:Adaptor Protein Complex 3; D033983:Adaptor Protein Complex mu Subunits; D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism* / Q000502:physiology; D002478:Cells, Cultured; D002966:Clathrin / Q000378:metabolism* / Q000502:physiology; D000089622:Doublecortin Domain Proteins; D004622:Embryo, Mammalian; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D006801:Humans; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000378:metabolism* / Q000502:physiology; D051379:Mice; D008869:Microtubule-Associated Proteins; D009420:Nervous System / Q000196:embryology* / Q000378:metabolism*; D009479:Neuropeptides / Q000378:metabolism* / Q000502:physiology; D010446:Peptide Fragments / Q000378:metabolism / Q000502:physiology; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D018808:Transcription Factor AP-1 / Q000378:metabolism
C438883:AP1M1 protein, human; C499064:AP1M2 protein, human; C510070:AP3M2 protein, human; C438890:APM1 protein, S cerevisiae; D033961:Adaptor Protein Complex 1; D033962:Adaptor Protein Complex 2; D033963:Adaptor Protein Complex 3; D033983:Adaptor Protein Complex mu Subunits; D033942:Adaptor Proteins, Vesicular Transport; C499063:Ap1m1 protein, mouse; C438884:Ap1m2 protein, mouse; C510071:Ap3m2 protein, mouse; D002352:Carrier Proteins; D002966:Clathrin; D000089622:Doublecortin Domain Proteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D009479:Neuropeptides; D010446:Peptide Fragments; D029701:Saccharomyces cerevisiae Proteins; D018808:Transcription Factor AP-1; C499066:adaptor protein complex 1, mu 1 subunit; C499065:adaptor protein complex 1, mu 2 subunit; C438885:adaptor protein complex 2, mu 1 subunit; C438886:adaptor protein complex 2, mu 2 subunit