The microtubule-associated protein doublecortin-like regulates the transport of the glucocorticoid receptor in neuronal progenitor cells.
In neuronal cells, activated glucocorticoid receptor (GR) translocates to the nucleus guided by the cytoskeleton. However, the detailed mechanisms underlying GR translocation remain unclear. Using gain and loss of function studies, we report here for the first time that the microtubule-associated protein doublecortin-like (DCL) controls GR translocation to the nucleus. DCL overexpression in COS-1 cells, neuroblastoma cells, and rat hippocampus organotypic slice cultures impaired GR translocation and decreased GR-dependent transcriptional activity, measured by a specific reporter gene assay, in COS-1 cells. Moreover, DCL and GR directly interact on microtubule bundles formed by DCL overexpression. A C-terminal truncated DCL with conserved microtubule-bundling activity did not influence GR translocation. In N1E-115 mouse neuroblastoma cells and neuronal progenitor cells in rat hippocampus organotypic slice cultures, laser-scanning confocal microscopy showed colabeling of endogenously expressed DCL and GR. In these systems, RNA-interference-mediated DCL knockdown hampered GR translocation. Thus, we conclude that DCL expression is tightly regulated to adequately control GR transport. Because DCL is primarily expressed in neuronal progenitor cells, our results introduce this microtubule-associated protein as a new modulator of GR signaling in this cell type and suggest the existence of cell-specific mechanisms regulating GR translocation to the nucleus.
Fitzsimons
Carlos P
CP
Leiden/Amsterdam Center for Drug Research/Medical Pharmacology Department, Einsteinweg 55, Leiden, The Netherlands.
Ahmed
Suaad
S
Wittevrongel
Christiaan F W
CF
Schouten
Theo G
TG
Dijkmans
Thomas F
TF
Scheenen
Wim J J M
WJ
Schaaf
Marcel J M
MJ
de Kloet
E Ronald
ER
Vreugdenhil
Erno
E
D000818:Animals; D001692:Biological Transport; D015153:Blotting, Western; D019556:COS Cells; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D000089662:Doublecortin Protein; D031541:Fluorescence Resonance Energy Transfer; D006624:Hippocampus / Q000378:metabolism; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D009154:Mutation; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
C000714848:Dcx protein, rat; D000089662:Doublecortin Protein; D008869:Microtubule-Associated Proteins; D011965:Receptors, Glucocorticoid