Multiple pathways regulate intracellular shuttling of MoKA, a co-activator of transcription factor KLF7.
MoKA is a novel F-box containing protein that interacts with and stimulates the activity of transcription factor KLF7, a regulator of neuronal differentiation. MoKA accumulates throughout the cell and predominantly in the cytosol, consistent with the presence of several putative nuclear localization and export signals (NLSs and NESs). The present study was designed to refine the identity and location of the sequences responsible for MoKA intracellular shuttling and transcriptional activity. Forced expression of fusion proteins in mammalian cells demonstrated that only one of three putative NLSs potentially recognized by karyopherin receptors is involved in nuclear localization of MoKA. By contrast, three distinct sequences were found to participate in mediating cytoplasmic accumulation. One of them is structurally and functionally related to the leucine-rich export signal that interacts with the exportin 1 (CRM1) receptor. The other two export signals instead display either a novel leucine-rich sequence or an undefined peptide motif, and both appear to act through CRM1-independent pathways. Finally, transcriptional analyses using the chimeric GAL4 system mapped the major activation domain of MoKA to a highly acidic sequence that resides between the NLS and NES clusters.
Smaldone
Silvia
S
Child Health Institute of New Jersey, Robert W. Johnson Medical School, 89 French Street, New Brunswick, NJ 08901, USA.
Ramirez
Francesco
F
D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002467:Cell Nucleus / Q000737:chemistry; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000737:chemistry; D044783:F-Box Proteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D028884:Karyopherins / Q000378:metabolism; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells; D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals; D021381:Protein Transport; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D015534:Trans-Activators / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D000097606:Exportin 1 Protein
D044783:F-Box Proteins; D028884:Karyopherins; D051741:Kruppel-Like Transcription Factors; D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals; D018160:Receptors, Cytoplasmic and Nuclear; D015534:Trans-Activators; D000097606:Exportin 1 Protein; C481794:Fbxo38 protein, mouse; C426008:Klf7 protein, mouse