Effect of Protein Kinase A Activity on the Association of ADP-ribosylation Factor 1 to Golgi Membranes

Pretreatment of alveolar macrophages with cholera toxin inhibits the release of arachidonic acid induced by the chemotactic peptide N-formylmethionyl-leucyl-phenylalanine. The results presented here show that cholera toxin might exert its inhibitory effect through the phosphorylation of Gi alpha by protein kinase A (PKA). (1) Gi-proteins from cells pretreated with cholera toxin showed parallel increases in their sensitivity to ADP-ribosylation by toxins in vitro and in Gi alpha phosphorylation. By contrast, the Gi alpha concentration was unchanged. (2) Cholera toxin pretreatment also decreased the functional activity of Gi, as assessed by the inhibition (80%) of agonist-induced binding of guanosine-5′-[gamma-thio]triphosphate (GTP[gamma S]). (3) These effects of cholera toxin were blocked by a specific PKA inhibitor, N-(2-[methyl-amino]ethyl)-3-isoquinolinesulphonamide dihydrochloride (H8) and mimicked by a cyclic AMP (cAMP) analogue and a phosphatase inhibitor. (4) Gi alpha was also phosphorylated in vitro by the catalytic subunit of PKA. In contrast with other cell systems, the stimulation of protein kinase C seems to have no effect on the sensitivity of Gi to ADP-ribosylation or on its phosphorylation. Therefore, the phosphorylation of Gi-proteins by PKA seems to be the actual target of the negative control of arachidonic acid release via the cAMP-mediated pathway.

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ADP-ribosylation of Rho proteins by Clostridium botulinum exoenzyme C3 is influenced by phosphorylation of Rho-associated factors

Biochemical Journal ◽

10.1042/bj3000133 ◽

1994 ◽

Vol 300 (1) ◽

pp. 133-139 ◽

Cited By ~ 13

Author(s):

G Fritz ◽

K Aktories

Keyword(s):

Alkaline Phosphatase ◽

Protein Kinase ◽

Fusion Protein ◽

Protein Kinase A ◽

Clostridium Botulinum ◽

Rho Proteins ◽

Cho Cell ◽

Adp Ribosylation ◽

Cell Extracts ◽

Kinase A

Specific [32P]ADP-ribosylation by Clostridium botulinum exoenzyme C3 was used to study the involvement of phosphorylation in the regulation of the low-molecular-mass GTP-binding protein Rho. Dephosphorylation of CHO cell extracts by alkaline phosphatase treatment resulted in a 80-90% reduction in the C3-catalysed [32P]ADP-ribosylation of Rho proteins in both cytosolic and membrane fractions. Similar results were obtained after dephosphorylation with protein phosphatase type-1 from bovine retina, whereas type-2B and type-2C phosphatases had no effect on the level of subsequent [32P]ADP-ribosylation of Rho by C3. Incubation of CHO cell lysate under phosphorylation conditions increased the subsequent C3-mediated [32P]ADP-ribosylation of Rho proteins. The protein kinase inhibitors H7 and H9 had no effect on [32P]ADP-ribosylation at concentrations which are specific for inhibition of protein kinase A or C. Recombinant glutathione S-transferase-RhoA fusion protein (GST-RhoA) was phosphorylated by protein kinase A; however, the phosphorylation had no stimulatory effect on the ADP-ribosylation of GST-RhoA by C3. An approx. 48 kDa phosphoprotein was identified which bound specifically to recombinant GST-RhoA fusion protein. By gel-permeation chromatography, Rho-containing complexes of approx. 50 kDa and 130-170 kDa were detected. The ADP-ribosylation of Rho in the 130-170 kDa complex was reduced by alkaline phosphatase pretreatment. The data suggest that Rho activity is influenced by phosphorylation of Rho-associated regulatory factors. Phosphorylation/dephosphorylation of these Rho-regulating factors appears to alter the ability of Rho to serve as a substrate for C3-induced [32P]ADP-ribosylation.

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cAMP has a protein kinase A independent regulatory effect on T cell proliferation, MAP kinase activity and on mRNA levels for IL-2 and INF-gamma

Immunology Letters ◽

10.1016/s0165-2478(97)87084-8 ◽

1997 ◽

Vol 56 (1-3) ◽

pp. 65

Author(s):

B Hofmann

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Protein Kinase ◽

Map Kinase ◽

Protein Kinase A ◽

Kinase Activity ◽

T Cell Proliferation ◽

Mrna Levels ◽

Regulatory Effect ◽

Kinase A

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2068 Human cardiac inward rectifier Kir2.2/Kir2.1b (KCNJ12) potassium channels are antagonistically regulated by protein kinase A and protein kinase C

European Heart Journal ◽

10.1016/s0195-668x(03)95062-7 ◽

2003 ◽

Vol 24 (5) ◽

pp. 382

Author(s):

S KATHOFER

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Potassium Channels ◽

Protein Kinase A ◽

Inward Rectifier ◽

Kinase C ◽

Kinase A

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Protein kinase D2-regulated protein transport at the trans-Golgi network requires its targeting to this compartment by ADP-ribosylation factor 1

Zeitschrift für Gastroenterologie ◽

10.1055/s-0030-1263799 ◽

2010 ◽

Vol 48 (08) ◽

Author(s):

GV Pusapati ◽

G Adler ◽

T Seufferlein

Keyword(s):

Protein Kinase ◽

Protein Transport ◽

Adp Ribosylation ◽

Trans Golgi Network ◽

Golgi Network ◽

Adp Ribosylation Factor

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Expression and mutation analysis of regulatory subunit Iα of protein kinase A in undifferentiated and anaplastic thyroid carcinoma

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/s-2004-819071 ◽

2004 ◽

Vol 112 (S 1) ◽

Author(s):

M Lippert ◽

SY Sheu ◽

K Worm ◽

M Wichert ◽

KW Schmid ◽

...

Keyword(s):

Protein Kinase ◽

Thyroid Carcinoma ◽

Protein Kinase A ◽

Mutation Analysis ◽

Anaplastic Thyroid Carcinoma ◽

Regulatory Subunit ◽

Kinase A

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cAMP exerts proliferative and anti-proliferative effects in pituitary cells of different types by activating both cAMP-dependent protein kinase A and exchange proteins directly activated by cAMP

Endocrine Abstracts ◽

10.1530/endoabs.32.p827 ◽

2013 ◽

Author(s):

Eleonora Vitali ◽

Erika Peverelli ◽

Giovanna Mantovani ◽

Elena Giardino ◽

Andrea G. Lania ◽

...

Keyword(s):

Protein Kinase ◽

Protein Kinase A ◽

Pituitary Cells ◽

Dependent Protein Kinase ◽

Camp Dependent Protein Kinase ◽

Different Types ◽

Dependent Protein ◽

Kinase A

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Regulation of spontaneous and induced resumption of meiosis in mouse oocytes by different intracellular pathways

Reproduction ◽

10.1530/reprod/120.2.377 ◽

2000 ◽

pp. 377-383 ◽

Cited By ~ 13

Author(s):

L Leonardsen ◽

A Wiersma ◽

M Baltsen ◽

AG Byskov ◽

CY Andersen

Keyword(s):

Signal Transduction ◽

Protein Kinase ◽

Protein Kinase A ◽

Mitogen Activated Protein Kinase ◽

Meiotic Maturation ◽

Mouse Oocytes ◽

Mitogen Activated Protein ◽

Dependent Signal ◽

Kinase Pathway ◽

Kinase A

The mitogen-activated protein kinase-dependent and the cAMP-protein kinase A-dependent signal transduction pathways were studied in cultured mouse oocytes during induced and spontaneous meiotic maturation. The role of the mitogen-activated protein kinase pathway was assessed using PD98059, which specifically inhibits mitogen-activated protein kinase 1 and 2 (that is, MEK1 and MEK2), which activates mitogen-activated protein kinase. The cAMP-dependent protein kinase was studied by treating oocytes with the protein kinase A inhibitor rp-cAMP. Inhibition of the mitogen-activated protein kinase pathway by PD98059 (25 micromol l(-1)) selectively inhibited the stimulatory effect on meiotic maturation by FSH and meiosis-activating sterol (that is, 4,4-dimethyl-5alpha-cholest-8,14, 24-triene-3beta-ol) in the presence of 4 mmol hypoxanthine l(-1), whereas spontaneous maturation in the absence of hypoxanthine was unaffected. This finding indicates that different signal transduction mechanisms are involved in induced and spontaneous maturation. The protein kinase A inhibitor rp-cAMP induced meiotic maturation in the presence of 4 mmol hypoxanthine l(-1), an effect that was additive to the maturation-promoting effect of FSH and meiosis-activating sterol, indicating that induced maturation also uses the cAMP-protein kinase A-dependent signal transduction pathway. In conclusion, induced and spontaneous maturation of mouse oocytes appear to use different signal transduction pathways.

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