title: Nuclear transport of Kir/Gem requires specific signals and importin alpha5 and is regulated by calmodulin and predicted serine phosphorylations.
abstract: Kir/Gem, together with Rad, Rem and Rem2, is a member of the RGK small GTP-binding protein family. These multifunctional proteins regulate voltage-gated calcium channel (VGCC) activity and cell-shape remodeling. Calmodulin and 14-3-3 binding modulate the functions of RGK proteins. Intriguingly, abolishing the binding of calmodulin or calmodulin and 14-3-3 results in nuclear accumulation of RGK proteins. Under certain conditions, the Ca(v)beta3-subunit of VGCCs can be translocated into the nucleus along with the RGK proteins, resulting in channel inactivation. The mechanism by which nuclear localization of RGK proteins is accomplished and regulated, however, is unknown. Here, we identify specific nuclear localization signals (NLS) in Kir/Gem that are both required and sufficient for nuclear transport. Importin alpha5 binds to Kir/Gem, and its depletion using RNA interference impairs nuclear translocation of this RGK protein. Calmodulin and predicted phosphorylations on serine residues within or in the vicinity of a C-terminal bipartite NLS regulate nuclear translocation by interfering with the association between importinalpha5 and Kir/Gem. These predicted phosphorylations, however, do not affect Kir/Gem-mediated calcium channel downregulation but rather, as shown in the accompanying paper (Mahalakshmi RN, Ng MY, Guo K, Qi Z, Hunziker W, Béguin P. Nuclear localization of endogenous RGK proteins and modulation of cell shape remodeling by regulated nuclear transport. Traffic 2007; doi:10.1111/j.1600-0854.2007.00599.x), interfere with cell-shape remodeling.
authors.affiliation: Epithelial Cell Biology Laboratory, Institute of Molecular and Cell Biology, 61 Biopolis Drive, Singapore 138673, Singapore.
authors.forename: Ramasubbu N
authors.initials: RN
authors.lastname: Mahalakshmi
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Nagashima
authors.forename: Mei Yong
authors.initials: MY
authors.lastname: Ng
authors.forename: Nobuya
authors.initials: N
authors.lastname: Inagaki
authors.forename: Walter
authors.initials: W
authors.lastname: Hunziker
authors.forename: Pascal
authors.initials: P
authors.lastname: Béguin
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D019556:COS Cells; D015220:Calcium Channels / Q000378:metabolism / Q000502:physiology; D002147:Calmodulin / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004594:Electrophysiology; D006367:HeLa Cells; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D028884:Karyopherins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D019913:Nuclear Localization Signals / Q000235:genetics / Q000502:physiology; D016716:PC12 Cells; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D051381:Rats; D012694:Serine / Q000378:metabolism*; D028901:alpha Karyopherins / Q000235:genetics / Q000378:metabolism*
chemical_list: D015220:Calcium Channels; D002147:Calmodulin; D017874:Immediate-Early Proteins; C423142:KPNA1 protein, human; D028884:Karyopherins; D019913:Nuclear Localization Signals; D028901:alpha Karyopherins; D012694:Serine; C088119:Gem protein, mouse; D020559:Monomeric GTP-Binding Proteins