scholarly journals Calcium/calmodulin-dependent protein kinase types II and IV differentially regulate CREB-dependent gene expression.

1994 ◽  
Vol 14 (9) ◽  
pp. 6107-6116 ◽  
Author(s):  
R P Matthews ◽  
C R Guthrie ◽  
L M Wailes ◽  
X Zhao ◽  
A R Means ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Cultured Cells ◽  
Active Form ◽  
Cell Types ◽  
Gene Induction ◽  
Lymphoid Tissues ◽  
Dependent Protein Kinase ◽  
Creb Phosphorylation ◽  
Dependent Protein ◽  
Constitutively Active

Phosphorylation of CREB (cyclic AMP [cAMP]- response element [CRE]-binding protein) by cAMP-dependent protein kinase (PKA) leads to the activation of many promoters containing CREs. In neurons and other cell types, CREB phosphorylation and activation of CRE-containing promoters can occur in response to elevated intracellular Ca2+. In cultured cells that normally lack this Ca2+ responsiveness, we confer Ca(2+)-mediated activation of a CRE-containing promoter by introducing an expression vector for Ca2+/calmodulin-dependent protein kinase type IV (CaMKIV). Activation could also be mediated directly by a constitutively active form of CaMKIV which is Ca2+ independent. The CaMKIV-mediated gene induction requires the activity of CREB/ATF family members but is independent of PKA activity. In contrast, transient expression of either a constitutively active or wild-type Ca2+/calmodulin-dependent protein kinase type II (CaMKII) fails to mediate the transactivation of the same CRE-containing reporter gene. Examination of the subcellular distribution of transiently expressed CaMKIV and CaMKII reveals that only CaMKIV enters the nucleus. Our results demonstrate that CaMKIV, which is expressed in neuronal, reproductive, and lymphoid tissues, may act as a mediator of Ca(2+)-dependent gene induction.

Calcium/calmodulin-dependent protein kinase types II and IV differentially regulate CREB-dependent gene expression

1994 ◽  
Vol 14 (9) ◽  
pp. 6107-6116
Author(s):  
R P Matthews ◽  
C R Guthrie ◽  
L M Wailes ◽  
X Zhao ◽  
A R Means ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Cultured Cells ◽  
Active Form ◽  
Cell Types ◽  
Gene Induction ◽  
Lymphoid Tissues ◽  
Dependent Protein Kinase ◽  
Creb Phosphorylation ◽  
Dependent Protein ◽  
Constitutively Active

Phosphorylation of CREB (cyclic AMP [cAMP]- response element [CRE]-binding protein) by cAMP-dependent protein kinase (PKA) leads to the activation of many promoters containing CREs. In neurons and other cell types, CREB phosphorylation and activation of CRE-containing promoters can occur in response to elevated intracellular Ca2+. In cultured cells that normally lack this Ca2+ responsiveness, we confer Ca(2+)-mediated activation of a CRE-containing promoter by introducing an expression vector for Ca2+/calmodulin-dependent protein kinase type IV (CaMKIV). Activation could also be mediated directly by a constitutively active form of CaMKIV which is Ca2+ independent. The CaMKIV-mediated gene induction requires the activity of CREB/ATF family members but is independent of PKA activity. In contrast, transient expression of either a constitutively active or wild-type Ca2+/calmodulin-dependent protein kinase type II (CaMKII) fails to mediate the transactivation of the same CRE-containing reporter gene. Examination of the subcellular distribution of transiently expressed CaMKIV and CaMKII reveals that only CaMKIV enters the nucleus. Our results demonstrate that CaMKIV, which is expressed in neuronal, reproductive, and lymphoid tissues, may act as a mediator of Ca(2+)-dependent gene induction.

scholarly journals Targeting of calcium/calmodulin-dependent protein kinase II

2004 ◽  
Vol 378 (1) ◽  
pp. 1-16 ◽  
Author(s):  
Roger J. COLBRAN
Keyword(s):  
Protein Kinase ◽  
Active Form ◽  
Cell Types ◽  
Dependent Protein Kinase ◽  
Calmodulin Binding ◽  
Calcium Calmodulin Dependent ◽  
Neuronal Dendrites ◽  
Protein Kinase Ii ◽  
Dependent Protein ◽  
Camkii Activation

Calcium/calmodulin-dependent protein kinase II (CaMKII) has diverse roles in virtually all cell types and it is regulated by a plethora of mechanisms. Local changes in Ca2+ concentration drive calmodulin binding and CaMKII activation. Activity is controlled further by autophosphorylation at multiple sites, which can generate an autonomously active form of the kinase (Thr286) or can block Ca2+/calmodulin binding (Thr305/306). The regulated actions of protein phosphatases at these sites also modulate downstream signalling from CaMKII. In addition, CaMKII targeting to specific subcellular microdomains appears to be necessary to account for the known signalling specificity, and targeting is regulated by Ca2+/calmodulin and autophosphorylation. The present review focuses on recent studies revealing the diversity of CaMKII interactions with proteins localized to neuronal dendrites. Interactions with various subunits of the NMDA (N-methyl-d-aspartate) subtype of glutamate receptor have attracted the most attention, but binding of CaMKII to cytoskeletal and several other regulatory proteins has also been reported. Recent reports describing the molecular basis of each interaction and their potential role in the normal regulation of synaptic transmission and in pathological situations are discussed. These studies have revealed fundamental regulatory mechanisms that are probably important for controlling CaMKII functions in many cell types.

cAMP-dependent protein kinase differentially regulates prestalk and prespore differentiation during Dictyostelium development

Development ◽  
1993 ◽  
Vol 119 (1) ◽  
pp. 135-146 ◽  
Author(s):  
S.K. Mann ◽  
R.A. Firtel
Keyword(s):  
Protein Kinase ◽  
Spatial Localization ◽  
Cell Types ◽  
Basal Region ◽  
Cell Type ◽  
Dependent Protein Kinase ◽  
Multicellular Development ◽  
Prespore Cell ◽  
Dependent Protein ◽  
Cell Type Specific

We and others have previously shown that cAMP-dependent protein kinase (PKA) activity is essential for aggregation, induction of prespore gene expression and multicellular development in Dictyostelium. In this manuscript, we further examine this regulatory role. We have overexpressed the Dictyostelium PKA catalytic subunit (PKAcat) in specific cell types during the multicellular stages, using prestalk and prespore cell-type-specific promoters to make PKA activity constitutive in these cells (independent of cAMP concentration). To examine the effects on cell-type differentiation, we cotransformed the PKAcat-expressing vectors with reporter constructs expressing lacZ from four cell-type-specific promoters: ecmA (specific for prestalk A cells); ecmB (specific for prestalk B and anterior-like cells in the slug); ecmB delta 89 (specific for stalk cells); and SP60 (prespore-cell-specific). By staining for beta-galactosidase expression histologically at various stages of development in individual strains, we were able to dissect the morphological changes in these strains, examine the spatial localization of the individual cell types, and understand the possible roles of PKA during multicellular development. Expression of PKAcat from either the ecmA or ecmB prestalk promoters resulted in abnormal development that arrested shortly after the mound stage, producing a mound with a round apical protrusion at the time of tip formation. Prestalk A and prestalk B cells were localized in the central region and the apical mound in the terminal differentiated aggregate, while prespore cells showed an aberrant spatial localization. Consistent with a developmental arrest, these mounds did not form either mature spores or stalk cells and very few cells expressed a stalk-cell-specific marker. Expression of PKAcat from the prespore promoter resulted in abnormal morphogenesis and accelerated spore cell differentiation. When cells were plated on agar, a fruiting body was formed with a very large basal region, containing predominantly spores, and a small, abnormal sorocarp. Mature spore cells were first detected by 14 hours, with maximal levels reached by 18–20 hours, in contrast to 24–26 hours in wild-type strains. When cells were plated on filters, they produced an elongated tip from a large basal region, which continued to elongate as a tubular structure and produce a ‘slug-like’ structure at the end. The slug was composed predominantly of prestalk cells with a few prespore cells restricted to the junction between the ‘slug’ and tube. As the slug migrated, these prespore cells were found in the tube, while new prespore cells appeared at the slug/tube junction, suggesting a continual differentiation of new prespore cells at the slug's posterior.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals An Active C-Terminally Truncated Form of Ca2+/Calmodulin-Dependent Protein Kinase Phosphatase-N (CaMKP-N/PPM1E)

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Atsuhiko Ishida ◽  
Kumiko Tsumura ◽  
Megu Oue ◽  
Yasuhiro Takenaka ◽  
Yasushi Shigeri ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Expression System ◽  
Active Form ◽  
Specific Inhibitor ◽  
Proteolytic Processing ◽  
Disulfonic Acid ◽  
Truncated Form ◽  
Dependent Protein Kinase ◽  
Wheat Embryo ◽  
Dependent Protein

Ca2+/calmodulin-dependent protein kinase phosphatase (CaMKP/PPM1F) and its nuclear homolog CaMKP-N (PPM1E) are Ser/Thr protein phosphatases that belong to the PPM family. CaMKP-N is expressed in the brain and undergoes proteolytic processing to yield a C-terminally truncated form. The physiological significance of this processing, however, is not fully understood. Using a wheat-embryo cell-free protein expression system, we prepared human CaMKP-N (hCaMKP-N(WT)) and the truncated form, hCaMKP-N(1–559), to compare their enzymatic properties using a phosphopeptide substrate. The hCaMKP-N(1–559) exhibited a much higherVmaxvalue than the hCaMKP-N(WT) did, suggesting that the processing may be a regulatory mechanism to generate a more active species. The active form, hCaMKP-N(1–559), showed Mn2+or Mg2+-dependent phosphatase activity with a strong preference for phospho-Thr residues and was severely inhibited by NaF, but not by okadaic acid, calyculin A, or 1-amino-8-naphthol-2,4-disulfonic acid, a specific inhibitor of CaMKP. It could bind to postsynaptic density and dephosphorylate the autophosphorylated Ca2+/calmodulin-dependent protein kinase II. Furthermore, it was inactivated by H2O2treatment, and the inactivation was completely reversed by treatment with DTT, implying that this process is reversibly regulated by oxidation/reduction. The truncated CaMKP-N may play an important physiological role in neuronal cells.

Activation of distinct cAMP- and cGMP-dependent pathways by relaxant agents in isolated gastric muscle cells

1993 ◽  
Vol 264 (3) ◽  
pp. G470-G477 ◽  
Author(s):  
J. G. Jin ◽  
K. S. Murthy ◽  
J. R. Grider ◽  
G. M. Makhlouf
Keyword(s):  
Protein Kinase ◽  
Muscle Cells ◽  
Cell Types ◽  
No Production ◽  
Taenia Coli ◽  
Dependent Protein Kinase ◽  
Cyclic Monophosphate ◽  
Cgmp Production ◽  
Gastric Muscle ◽  
Dependent Protein

The mechanism of action of vasoactive intestinal peptide (VIP) was examined in isolated gastric and taenia coli muscle cells and compared with that of nitric oxide (NO), sodium nitroprusside (SNP), and isoproterenol. In gastric muscle cells, VIP stimulated NO production, increased adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP) levels, and induced relaxation in a concentration-dependent fashion. The NO synthase inhibitor NG-nitro-L-arginine abolished NO and cGMP production and partly inhibited relaxation. The soluble guanylate cyclase inhibitor LY 83583 abolished cGMP production and partly inhibited relaxation. (R)-p-adenosine 3',5'-cyclic phosphorothioate [(R)-p-cAMPS], a preferential inhibitor of cAMP-dependent protein kinase (cAK), and KT5823, a preferential inhibitor of cGMP-dependent protein kinase (cGK), partly inhibited relaxation separately and abolished relaxation in combination. The pattern implied that VIP induced relaxation by activation of cAK and by NO-mediated stimulation of cGMP and activation of cGK. In taenia coli muscle cells, VIP did not increase NO production or cGMP levels: relaxation was accompanied by an increase in cAMP and was partly inhibited by (R)-p-cAMPS and KT5823 and abolished by a combination of both inhibitors. Isoproterenol increased only cAMP levels in both cell types, which induced relaxation by activating cAK at low concentrations of agonist and both cAK and cGK at high concentrations in a pattern identical to that observed with VIP in taenia coli muscle cells. SNP and NO increased only cGMP levels in both cell types, which induced relaxation by activating cGK only. We conclude that cAK and cGK can be activated separately and mediate relaxation independently.(ABSTRACT TRUNCATED AT 250 WORDS)

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