title: Constitutively elevated nuclear export activity opposes Ca2+-dependent NFATc3 nuclear accumulation in vascular smooth muscle: role of JNK2 and Crm-1.
abstract: The transcription factor NFAT (nuclear factor of activated T-cells) is a cytosolic phosphoprotein that accumulates in the nucleus following dephosphorylation by the calcium (Ca2+)/calmodulin-dependent phosphatase, calcineurin. A defining feature of stimuli that induce NFAT nuclear accumulation/activation is a sustained increase in global intracellular Ca2+. Contrary to expectations, we have found that a sustained elevation of intracellular Ca2+, induced by membrane potential depolarization and mediated by voltage-dependent Ca2+ channels, does not result in nuclear localization of the NFATc3 isoform in smooth muscle. However, vasoconstrictors (e.g. uridine triphosphate (UTP)) and growth factors, which elevate intracellular Ca2+ and engage multiple intracellular signaling pathways, induce a robust increase in smooth muscle nuclear NFATc3. Here we show that depolarizing stimuli that normally fail to induce NFATc3 nuclear accumulation in arterial smooth muscle effectively induce nuclear accumulation under conditions in which Crm-1-dependent or JNK2-mediated nuclear export processes are disrupted. Consistent with an important regulatory role for JNK, UTP exerts a suppressive effect on JNK activity in smooth muscle. Export of nuclear NFATc3 following UTP-induced nuclear accumulation is dramatically slowed in cerebral arteries from JNK2-/- animals. These data indicate that in smooth muscle, stimulation of Ca2+-dependent, calcineurin-mediated nuclear import and suppression of Crm-1/JNK-dependent nuclear export are both required for induction of NFATc3 nuclear accumulation. These results highlight the dynamic interplay between influences that promote and oppose NFAT nuclear accumulation and suggest that in arterial smooth muscle suppression of constitutive nuclear export activity is an important property of NFAT-activating stimuli.
authors.affiliation: Department of Pharmacology, University of Vermont, 89 Beaumont Avenue, Burlington, VT 05405, USA.
authors.forename: Maria F
authors.initials: MF
authors.lastname: Gomez
authors.forename: Laura V Gonzalez
authors.initials: LV
authors.lastname: Bosc
authors.forename: Andra S
authors.initials: AS
authors.lastname: Stevenson
authors.forename: M Keith
authors.initials: MK
authors.lastname: Wilkerson
authors.forename: David C
authors.initials: DC
authors.lastname: Hill-Eubanks
authors.forename: Mark T
authors.initials: MT
authors.lastname: Nelson
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D000818:Animals; D019703:Calcineurin / Q000502:physiology; D002118:Calcium / Q000494:pharmacology; D002536:Cerebral Arteries; D004268:DNA-Binding Proteins / Q000378:metabolism*; D028884:Karyopherins / Q000502:physiology; D008564:Membrane Potentials; D051379:Mice; D018345:Mice, Knockout; D048056:Mitogen-Activated Protein Kinase 9; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors / Q000378:metabolism*; D014544:Uridine Triphosphate / Q000494:pharmacology; D014662:Vasoconstrictor Agents / Q000494:pharmacology; D000097606:Exportin 1 Protein
chemical_list: D019703:Calcineurin; D002118:Calcium; D004268:DNA-Binding Proteins; D028884:Karyopherins; D048056:Mitogen-Activated Protein Kinase 9; D020928:Mitogen-Activated Protein Kinases; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors; D014544:Uridine Triphosphate; D014662:Vasoconstrictor Agents; D000097606:Exportin 1 Protein; C494009:Nfatc3 protein, mouse; C445503:transcription factor NF-AT c3