title: Featured Article: Nuclear export of opioid growth factor receptor is CRM1 dependent.
abstract: Opioid growth factor receptor (OGFr) facilitates growth inhibition in the presence of its specific ligand opioid growth factor (OGF), chemically termed [Met(5)]-enkephalin. The function of the OGF-OGFr axis requires the receptor to translocate to the nucleus. However, the mechanism of nuclear export of OGFr is unknown. In this study, endogenous OGFr, as well as exogenously expressed OGFr-EGFP, demonstrated significant nuclear accumulation in response to leptomycin B (LMB), an inhibitor of CRM1-dependent nuclear export, suggesting that OGFr is exported in a CRM1-dependent manner. One consensus sequence for a nuclear export signal (NES) was identified. Mutation of the associated leucines, L217 L220 L223 and L225, to alanine resulted in decreased nuclear accumulation. NES-EGFP responded to LMB, indicating that this sequence is capable of functioning as an export signal in isolation. To determine why the sequence functions differently in isolation than as a full length protein, the localization of subNES was evaluated in the presence and absence of MG132, a potent inhibitor of proteosomal degradation. MG132 had no effect of subNES localization. The role of tandem repeats located at the C-terminus of OGFr was examined for their role in nuclear trafficking. Six of seven tandem repeats were removed to form deltaTR. DeltaTR localized exclusively to the nucleus indicating that the tandem repeats may contribute to the localization of the receptor. Similar to the loss of cellular proliferation activity (i.e. inhibition) recorded with subNES, deltaTR also demonstrated a significant loss of inhibitory activity indicating that the repeats may be integral to receptor function. These experiments reveal that OGFr contains one functional NES, L217 L220 L223 and L225 and can be exported from the nucleus in a CRM1-dependent manner.
authors.affiliation: Department of Neural & Behavioral Sciences, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, PA, USA.
authors.forename: Nancy P
authors.initials: NP
authors.lastname: Kren
authors.affiliation: Department of Neural & Behavioral Sciences, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, PA, USA.
authors.forename: Ian S
authors.initials: IS
authors.lastname: Zagon
authors.affiliation: Department of Neural & Behavioral Sciences, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, PA, USA pxm9@psu.edu.
authors.forename: Patricia J
authors.initials: PJ
authors.lastname: McLaughlin
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D028884:Karyopherins / Q000378:metabolism*; D049790:Nuclear Export Signals; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D011957:Receptors, Opioid / Q000235:genetics / Q000378:metabolism*; D000097606:Exportin 1 Protein
chemical_list: D028884:Karyopherins; D049790:Nuclear Export Signals; D018160:Receptors, Cytoplasmic and Nuclear; D011957:Receptors, Opioid; D000097606:Exportin 1 Protein; C033593:methionine-enkephalin receptor
keywords: CRM1; OGFr; leptomycin B; nuclear export signaling