Calcium-dependent enhancement of calcium current in smooth muscle by calmodulin-dependent protein kinase II

Nature ◽  
1992 ◽  
Vol 357 (6373) ◽  
pp. 74-77 ◽  
Author(s):  
John G. McCarron ◽  
J. Graham McGeown ◽  
Sheila Reardon ◽  
Mrtsuo Ikebe ◽  
Fredric S. Fay ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase ◽  
Calcium Current ◽  
Dependent Protein Kinase ◽  
Calcium Dependent ◽  
Protein Kinase Ii ◽  
Dependent Protein

iNOS regulation by calcium/calmodulin-dependent protein kinase II in vascular smooth muscle

2007 ◽  
Vol 292 (6) ◽  
pp. H2634-H2642 ◽  
Author(s):  
Rachel J. Jones ◽  
David Jourd'heuil ◽  
John C. Salerno ◽  
Susan M. E. Smith ◽  
Harold A. Singer
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase ◽  
Vascular Smooth Muscle ◽  
Resting Cells ◽  
Dependent Manner ◽  
Dependent Protein Kinase ◽  
Cytokine Induction ◽  
Calcium Calmodulin Dependent ◽  
Protein Kinase Ii ◽  
Dependent Protein

Nitric oxide synthase (NOS) expression is regulated transcriptionally in response to cytokine induction and posttranslationally by palmitoylation and trafficking into perinuclear aggresome-like structures. We investigated the effects of multifunctional calcium/calmodulin-dependent protein kinase II protein kinase (CaMKII) on inducible NOS (iNOS) trafficking in cultured rat aortic vascular smooth muscle cells (VSMCs). Immunofluorescence and confocal microscopy demonstrated colocalization of iNOS and CaMKIIδ2 with a perinuclear distribution and concentration in aggresome-like structures identified by colocalization with γ-tubulin. Furthermore, CaMKIIδ2 coimmunoprecipitated with iNOS in a CaMKII activity-dependent manner. Addition of Ca2+-mobilizing stimuli expected to activate CaMKII; a purinergic agonist (UTP) or calcium ionophore (ionomycin) caused a general redistribution of iNOS from cytosolic to membrane and nuclear fractions. Similarly, adenoviral expression of a constitutively active CaMKIIδ2 mutant altered iNOS localization, shifting iNOS from the cytosolic fraction. Suppression of CaMKIIδ2 using an adenovirus expressing a short hairpin, small interfering RNA increased nuclear iNOS localization in resting cells but inhibited ionomycin-induced translocation of iNOS to the nucleus. Following addition of these chronic and acute CaMKII modulators, there were fewer aggresome-like structures containing iNOS. All of the treatments that chronically affected CaMKII activity or expression significantly inhibited iNOS-specific activity following cytokine induction. The results suggest that CaMKIIδ2 may be an important regulator of iNOS trafficking and activity in VSMCs.

Abstract 17: Nuclear Calcium/Calmodulin-dependent Protein Kinase II Signaling Enhances Cardiac Progenitor Cell Survival and Cardiac Lineage Commitment

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Pearl Quijada ◽  
Nirmala Hariharan ◽  
Jonathan Cubillo ◽  
Kristin M Bala ◽  
Lucia Ormachea ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase ◽  
Troponin T ◽  
Morphological Changes ◽  
Smooth Muscle Actin ◽  
Cardiac Differentiation ◽  
Dependent Protein Kinase ◽  
Nuclear Compartment ◽  
Protein Kinase Ii ◽  
Dependent Protein

Ca2+/Calmodulin-dependent protein kinase II (CaMKII) signaling in the heart regulates cardiomyocyte contractility and growth in response to elevated intracellular Ca2+. The δB isoform of CaMKII is the predominant nuclear splice variant in the adult heart and regulates cardiomyocyte hypertrophic gene expression by signaling to the histone deacetylase HDAC4. However, the role of CaMKIIδ in cardiac progenitor cells (CPCs) has not been explored. During developmental growth endogenous CPCs display primarily cytosolic CaMKIIδ, which localizes to the nuclear compartment of CPCs after myocardial infarction injury. CPCs undergoing early differentiation in vitro increase levels of CaMKIIδB in the nuclear compartment where the kinase may contribute to the regulation of CPC commitment. CPCs modified with an established lentiviral based constructs to overexpress CaMKIIδB (CPCeδB) have reduced proliferative rate compared to lentiviral transduction of CPCs with eGFP alone (CPCe). Additionally, stable expression of CaMKIIδB promotes distinct morphological changes such as increased cell surface area and increased length of cells compared to CPCe. CPCeδB are resistant to oxidative stress induced by H2O2 relative to CPCe, whereas a knockdown of CaMKIIδB using small hairpin RNA resulted in an up regulation of cell death compared to scrambled treated controls. Dexamethasone treatment to promote cardiac differentiation increased cardiomyogenic markers cardiac troponin T and α-smooth muscle actin measured by RT-PCR and immunoblot analyses in CPCeδB compared to control CPCe. Therefore, CaMKIIδB may serve as a novel modulatory protein to enhance CPC survival and commitment into the cardiac and smooth muscle lineage.

scholarly journals Calponin phosphorylation in vitro and in intact muscle

1993 ◽  
Vol 296 (3) ◽  
pp. 827-836 ◽  
Author(s):  
S J Winder ◽  
B G Allen ◽  
E D Fraser ◽  
H M Kang ◽  
G J Kargacin ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Gel Electrophoresis ◽  
Dependent Protein Kinase ◽  
Kinase C ◽  
Sds Page ◽  
Protein Kinase Ii ◽  
Dependent Protein

Calponin, a thin-filament-associated protein implicated in the regulation of smooth-muscle contraction, is phosphorylated in vitro by protein kinase C and Ca2+/calmodulin-dependent protein kinase II [Winder and Walsh (1990) J. Biol. Chem. 265, 10148-10155] and dephosphorylated by a type 2A protein phosphatase [Winder, Pato and Walsh (1992) Biochem. J. 286, 197-203]. Unphosphorylated calponin binds to actin and inhibits the actin-activated myosin MgATPase; these properties are lost on phosphorylation. Although both serine and threonine residues in calponin are phosphorylated, the major site of phosphorylation by either kinase is Ser-175. Calponin also undergoes phosphorylation when bound to actin in synthetic thin filaments, in a reconstituted actomyosin system, in washed myofibrils and in tissue extracts; this results in dissociation of calponin from actin. Tryptic phosphopeptide mapping indicates that the same sites are phosphorylated in the bound as in the isolated protein. Toad stomach calponin exists in at least three isoforms which differ in charge but exhibit the same molecular mass on SDS/PAGE. In a toad stomach extract, all three isoforms are phosphorylated by protein kinase C or Ca2+/calmodulin-dependent protein kinase II as shown by two-dimensional gel electrophoresis (non-equilibrium pH-gradient gel electrophoresis and SDS/PAGE). Calponin phosphorylation also occurs in intact toad stomach smooth-muscle strips metabolically labelled with 32Pi and stimulated to contract with carbachol. These results support the hypothesis that calponin may be regulated in vivo by phosphorylation-dephosphorylation.

Calcium/calmodulin-dependent protein kinase II-δ isoform regulation of vascular smooth muscle cell proliferation

2007 ◽  
Vol 292 (6) ◽  
pp. C2276-C2287 ◽  
Author(s):  
Suzanne J. House ◽  
Roman G. Ginnan ◽  
Shayn E. Armstrong ◽  
Harold A. Singer
Keyword(s):  
Cell Proliferation ◽  
Smooth Muscle ◽  
Protein Kinase ◽  
Vascular Smooth Muscle ◽  
Dna Synthesis ◽  
Primary Culture ◽  
Primary Cultures ◽  
Dependent Protein Kinase ◽  
Protein Kinase Ii ◽  
Dependent Protein

There is accumulating evidence that Ca2+-dependent signaling pathways regulate proliferation and migration of vascular smooth muscle (VSM) cells, contributing to the intimal accumulation of VSM that is a hallmark of many vascular diseases. In this study we investigated the role of the multifunctional serine/threonine kinase, calmodulin (CaM)-dependent protein kinase II (CaMKII), as a mediator of Ca2+ signals regulating VSM cell proliferation. Differentiated VSM cells acutely isolated from rat aortic media express primarily CaMKIIγ gene products, whereas passaged primary cultures of de-differentiated VSM cells express primarily CaMKIIδ2, a splice variant of the δ gene. Experiments examining the time course of CaMKII isoform modulation revealed the process was rapid in onset following initial dispersion and primary culture of aortic VSM with a significant increase in CaMKIIδ2 protein and a significant decrease in CaMKIIγ protein within 30 h, coinciding with the onset of DNA synthesis and cell proliferation. Attenuating the initial upregulation of CaMKIIδ2 in primary cultured cells using small-interfering RNA (siRNA) resulted in decreased serum-stimulated DNA synthesis and cell proliferation in primary culture. In passaged VSM cells, suppression of CaMKIIδ2 activity by overexpression of a kinase-negative mutant, or suppression of endogenous CaMKII content using multiple siRNAs, significantly attenuated serum-stimulated DNA synthesis and cell proliferation. Cell cycle analysis following either inhibitory approach indicated decreased proportion of cells in G1, an increase in proportion of cells in G2/M, and an increase in polyploidy, corresponding with accumulation of multinucleated cells. These results indicate that CaMKIIδ2 is specifically induced during modulation of VSM cells to the synthetic phenotypic and is a positive regulator of serum-stimulated proliferation.

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