okadaic-acid and oxophenylarsine

In the early secretory pathway, a distinct set of processing enzymes and family of lectins facilitate the folding and quality control of newly synthesized glycoproteins. In this regard, we recently identified a mechanism in which processing by endoplasmic reticulum mannosidase I, which attenuates the removal of glucose from asparagine-linked oligosaccharides, sorts terminally misfolded alpha(1)-antitrypsin for proteasome-mediated degradation in response to its abrogated physical dissociation from calnexin (Liu, Y., Choudhury, P., Cabral, C., and Sifers, R. N. (1999) J. Biol. Chem. 274, 5861-5867). In the present study, we examined the quality control of genetic variant PI Z, which undergoes inappropriate polymerization following biosynthesis. Here we show that in stably transfected hepatoma cells the additional processing of asparagine-linked oligosaccharides by endoplasmic reticulum mannosidase II partitions variant PI Z away from the conventional disposal mechanism in response to an arrested posttranslational interaction with calnexin. Intracellular disposal is accomplished by a nonproteasomal system that functions independently of cytosolic components but is sensitive to tyrosine phosphatase inhibition. The functional role of ER mannosidase II in glycoprotein quality control is discussed.

Topics: alpha 1-Antitrypsin; Animals; Arsenicals; Carbohydrate Sequence; Endoplasmic Reticulum; Genetic Variation; Glycoproteins; Humans; Liver Neoplasms, Experimental; Mannosidases; Mice; Molecular Sequence Data; Okadaic Acid; Oligosaccharides; Recombinant Proteins; Sodium Fluoride; Transfection; Tumor Cells, Cultured; Vanadates

In the capping of cell-surface receptors two stages can be distinguished: 1) clustering of the receptors (patching) induced by cross-linking with specific antibodies and 2) subsequent assembly of patches into a cap which is driven by the actin-based cytoskeleton. We found that patching of Fcgamma receptor II in U937 cells was correlated with tyrosine phosphorylation of certain proteins, most prominently those of 130, 110, 75 and 28 kDa. The phosphotyrosine-bearing proteins were accumulated at the receptor patches. Formation of the receptor caps was coincident with dephosphorylation of these proteins. Inhibition of protein tyrosine kinases with herbimycin A and genistein attenuated the protein tyrosine hyperphosphorylation and blocked capping in a dose-dependent manner. Phenylarsine oxide and pervanadate, inhibitors of protein tyrosine phosphatases, also suppressed capping of Fcgamma receptor II in a concentration-dependent fashion. Simultaneously, tyrosine hyperphosphorylation of proteins occurred. In the presence of the tyrosine kinase and phosphatase inhibitors the receptors were arrested at the patching stage. In contrast, okadaic acid, a serine/threonine phosphatase blocker, did not affect assembly of the receptor caps. The inhibitory effect of phenylarsine oxide was rapidly reversed by dithiols, 2,3-dimercapto-1-propanoldithiol and dithiotreitol, and was coincident with dephosphorylation of protein tyrosine residues. Extensive washing of pervanadate-exposed cells also resulted in progressive restoration of the cap assembly. Using streptolysin O-permeabilized cells we confirmed regulatory function played by dephosphorylation of tyrosine residues in capping of Fcgamma receptor II. Exogenous phosphatases, applied to permeabilized cells in which activity of endogenous tyrosine phosphatases was blocked, evoked dephosphorylation of protein tyrosine residues that was accompanied by recovery of capping ability in the cells.

Topics: Arsenicals; Bacterial Proteins; Benzoquinones; Cell Membrane Permeability; Dose-Response Relationship, Drug; Enzyme Inhibitors; Genistein; Humans; Immunoblotting; Lactams, Macrocyclic; Microscopy, Fluorescence; Okadaic Acid; Phosphorylation; Quinones; Receptor Aggregation; Receptors, IgG; Rifabutin; Streptolysins; Temperature; Time Factors; Tyrosine; U937 Cells; Vanadates

Since angiotensin II (Ang II)-induced receptor internalization is required to maintain the production of certain intracellular signals in some target cells, we investigated the relationships between Ang II receptor endocytosis and the generation of second messengers in rat hepatocytes. The results of the present study demonstrate that in response to exposure of hepatocytes to Ang II, a decrease in surface Ang II receptors occurred, consistent with a rapid endocytosis of the receptor-bound hormone complex. Pretreatment of cells with okadaic acid (OA) did not have any effect on receptor-mediated internalization. In contrast, a marked reduction of the Ang II receptor endocytosis process occurred after treatment of hepatocytes with phenylarsine oxide (PAO), indicating that cysteine residues could be involved in receptor-mediated endocytosis. Stimulation of cells with Ang II blocked the generation of cyclic adenosine monophosphate (cAMP), which follows the stimulation of hepatocytes with forskolin. Moreover, Ang II increased both inositol 4,5-bisphosphate (IP2) and inositol 1,4,5-trisphosphate (IP3) generation, and enhanced intracellular calcium concentration ([Ca2+]i). Exposure of cells to PAO did not alter the effect of Ang II on the accumulation of cAMP after forskolin stimulation, indicating that endocytosis of the agonist-receptor complex is not involved in adenylate cyclase inhibition. Conversely, PAO and OA markedly reduced IP2 and IP3 synthesis, and the plateau phase of Ang II-induced Ca2+ mobilization. The relationship between Ang II-induced endocytosis and the generation of phosphoinositols and increment in [Ca2+]i indicates that sequestration of the Ang II receptor is necessary to maintain the production of these intracellular signals in rat hepatocytes.

Topics: Angiotensin II; Animals; Arsenicals; Cells, Cultured; Endocytosis; Enzyme Inhibitors; Hepatocytes; Male; Okadaic Acid; Rats; Rats, Wistar; Receptors, Angiotensin; Second Messenger Systems; Signal Transduction; Time Factors

The contributions were determined in primary cultures of bovine corneal epithelial cells (BCEC) of Na:H exchange (NHE) and vacuolar H+-ATPase (i.e. V-type) activity to the regulation of intracellular pH (pHi). Furthermore, we characterized the effects on pHi regulation of exposure to 1 microM ET-1 under control and acid loaded conditions. With the pH sensitive dye, 2',7' Bis (carboxyethyl)-5,6-carboxyfluorescein acetoxymethyl ester (BCECF-AM), the control pHi was 7.1 in NaCl (nominally HCO3-free) Ringers. Inhibition of NHE with 100 microM dimethylamiloride (DMA) rapidly decreased pHi by 0.37 units. Similarly, selective inhibition of V-type H+-ATPase with 10 microM bafilomycin A1 decreased pHi by 0.22 units. Following acid loading in NaCl Ringers with a 20 mm NH4Cl prepulse, pHi recovery was partially inhibited by exposure to either Na-free (NMGCl) Ringers, 100 microM DMA or 20 microM bafilomycin A1. Based on decreases in H+ efflux resulting from selective inhibition of NHE and V-type H+ pump activity, NHE activity accounts for 76% of the pHi recovery following acid loading. Under control conditions, ET-1 (1 microM) had no effect on pHi whereas ET-1 completely suppressed pHi recovery following acid loading in NaCl or NMGCl Ringers. This inhibitory effect was largely due to stimulation of ETA because in the presence of BQ-123 (10 microM), a selective ETA receptor antagonist, pHi recovery was completely restored. Suppression of pHi recovery also occurred following stimulation of protein kinase C (PKC) with 10(-7) m phorbol myristate (PMA) whereas 10(-7) m 4 alpha phorbol 12,13 didecanoate (PDD) had no effect. ET-1 failed to suppress pHi recovery after inhibition of PKC with 0.5 microM calphostin C suggesting that the inhibition of pHi recovery by ET-1 is a consequence of PKC stimulation. Similarly, inhibition of Ca2+-dependent calmodulin stimulated CaM II kinase with KN-62 (10 microM) reversed the suppression of pHi recovery by ET-1. Preinhibition of either protein phosphatase (PP), PP-1, PP-2A or PP-2B activity with 1 microM phenylarsine oxide, 10 nm okadaic acid, 10 microM cyclosporin A1 or 20 microM BAPTA, also obviated the suppression of pHi recovery by ET-1. Therefore ETA receptor mediated inhibition of pHi regulation following acid loading could be a consequence of either PKC or CaMII kinase stimulation. Each one of these kinases may in turn phosphorylate and thereby stimulate the activities of PP-1, PP-2A or PP-2B. An increase in the activity of any o

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Amiloride; Animals; Arsenicals; Calcium-Calmodulin-Dependent Protein Kinases; Cattle; Cells, Cultured; Chelating Agents; Cyclosporine; Diuretics; Egtazic Acid; Endothelin Receptor Antagonists; Endothelin-1; Enzyme Inhibitors; Epithelium, Corneal; Hydrogen-Ion Concentration; Immunosuppressive Agents; Intracellular Fluid; Naphthalenes; Okadaic Acid; Peptides, Cyclic; Phosphoprotein Phosphatases; Protein Kinase C; Receptors, Endothelin; Spectrometry, Fluorescence; Tetradecanoylphorbol Acetate

Because melanoma tumors originate partly from excessive UV exposure but become relatively resistant to radiation, we have now compared the effects of okadaic acid, a phosphatase inhibitor, with that of the thymidine analog bromodeoxyuridine as sensitizers of DNA damage in B16 melanoma. We now show that 25 nM okadaic acid promotes DNA fragmentation in B16 melanoma, increasing cell detachment as well as pigmentation, a characteristic of melanocytic cell differentiation. At lower levels, okadaic acid synergizes with UV exposure to increase DNA fragmentation. Although bromodeoxyuridine also caused DNA damage, it did not increase pigmentation and it suppressed cell detachment. Okadaic acid was also more effective in promoting DNA laddering in growing versus quiescent melanocytes. Because DNA damaging effects of okadaic acid are mediated by different pathways from those used by nucleoside analogs, like bromodeoxyuridine, we propose their concurrent effect with radiation as sensitizers to DNA damage.

Topics: Animals; Antimetabolites, Antineoplastic; Arsenicals; Bromodeoxyuridine; Cell Adhesion; Cell Differentiation; Cycloheximide; DNA Damage; DNA Fragmentation; Enzyme Inhibitors; Melanoma; Mice; Okadaic Acid; Phosphoprotein Phosphatases; Pigmentation; Protein Synthesis Inhibitors; Radiation-Sensitizing Agents; Tumor Cells, Cultured; Ultraviolet Rays

Angiotensin II (Ang II) provokes rapid internalisation of its receptor from plasma membranes in isolated rat hepatocytes. After 10 min stimulation with Ang II, plasma membrane lost about 60% of its 125I-Ang II-binding capacity. Internalisation was blocked by phenylarsine oxide (PhAsO), whereas okadaic acid, which markedly reduced the sustained phase of calcium mobilization, did not have a preventive effect on Ang II-receptor complex sequestration. These data suggest that Ang II receptor internalisation is probably independent of a phosphorylation/dephosphorylation cycle of critical serine/threonine residues in the receptor molecule. To establish a relationship between sequestration of the Ang II receptor and the physical properties of the Ang II-binding sites, 125I-Ang II-receptor complex profiles were analysed by isoelectric focusing. In plasma membrane preparations two predominant Ang II-binding sites, migrating to pI 6.8 and 6.5, were found. After exposure to Ang II, cells lost 125I-Ang II-binding capacity to the Ang II-receptor complex migrating at pI 6.8 which was prevented in PhAsO-treated cells. Pretreatment of hepatocytes with okadaic acid did not modify Ang II-receptor complex profiles, indicating that the binding sites corresponding to pI 6.5 and pI 6.8 do not represent a phosphorylated and/or non-phosphorylated form of the Ang II receptor. The results show that the Ang II-receptor complex isoform at pI 6.8 represents a functional form of the type-1 Ang II receptor. Further studies are necessary to identify the Ang II-related nature of the binding sites corresponding to pI 6.5.

Topics: Angiotensin II; Animals; Arsenicals; Binding Sites; Calcium; Cell Membrane; Cells, Cultured; Enzyme Inhibitors; Ions; Isoelectric Focusing; Isomerism; Liver; Male; Okadaic Acid; Protein Binding; Rats; Rats, Wistar; Receptors, Angiotensin

Vanadate mimics, whereas phenylarsine oxide (PAO) antagonizes, the effects of insulin in rat adipocytes. Both vanadate and PAO are documented inhibitors of protein-phosphotyrosine phosphatases. The relationship between the inhibition of 'inhibitory' PTPase and 'stimulatory' PTPase has been studied here in primary rat adipocytes. Low concentrations of PAO (IC50 = 0.6-2.0 microM) blocked the stimulating effects of insulin, vanadate and pervanadate on hexose uptake and glucose metabolism. Inhibition of isoproterenol-mediating lipolysis by vanadate and insulin was not blocked by PAO. The activating effects of okadaic acid on hexose uptake and glucose metabolism, which occur at points downstream to tyrosine phosphorylation, were also not blocked by PAO. Subsequent studies suggested that the PAO-sensitive PTPase comprises a minute fraction of the total adipocytic PTPase activity. To identify its location we applied procedures involving fractionations and activation of non-receptor adipocytic protein tyrosine kinase by PAO and vanadate in cell free assays. We found that the 'inhibitory' PTPase is exclusively associated with the membrane fraction whereas the 'stimulatory' PTPases are present in both the cytosolic and plasma membrane compartments. We next searched for markers, possibly associated with PAO-dependent desensitization and found that several proteins became phosphorylated on tyrosine moieties in the supernatant of PAO but not in vanadate pretreated adipocytes. In summary, we propose the presence of a minute, plasma membrane associated PTPase in primary rat adipocytes, inhibition of which arrests the activation of glucose metabolism. In contrast, inhibition of all the other cellular adipose PTPases, ultimately activates rather than inhibits these same bioeffects.

Topics: 3-O-Methylglucose; Adipocytes; Animals; Arsenicals; Cells, Cultured; Enzyme Inhibitors; Epididymis; Ethers, Cyclic; Glycerol; Glycolysis; Insulin; Isoproterenol; Kinetics; Lipolysis; Male; Methylglucosides; Okadaic Acid; Protein Tyrosine Phosphatases; Protein-Tyrosine Kinases; Rats; Rats, Wistar; Vanadates

During germination of barley grains, DNA fragmentation was observed in the aleurone. The appearance of DNA fragmentation in the aleurone layer, observed by TUNEL staining in aleurone sections, started near the embryo and extended to the aleurone cells far from the embryo in a time dependent manner. The same spatial temporal activities of hydrolytic enzymes such as alpha-amylase were observed in aleurone. DNA fragmentation could also be seen in vitro under osmotic stress, in isolated aleurone. During aleurone protoplast isolation, a very enhanced and strong DNA fragmentation occurred which was not seen in protoplast preparations of tobacco leaves. ABA was found to inhibit DNA fragmentation occurring in barley aleurone under osmotic stress condition and during protoplast isolation, while the plant growth regulator gibberellic acid counteracted the effect of ABA. Addition of auxin or cytokinin had no significant effect on DNA fragmentation in these cells. To study the role of phosphorylation in ABA signal transduction leading to control of DNA fragmentation (apoptosis), the effects of the phosphatase inhibitor okadaic acid and of phenylarisine oxide on apoptosis were studied. We hypothesize that the regulation of DNA fragmentation in aleurone plays a very important role in spatial and temporal control of aleurone activities during germination. The possible signal transduction pathway of ABA leading to the regulation of DNA fragmentation is discussed.

Topics: Abscisic Acid; Apoptosis; Arsenicals; DNA Fragmentation; Enzyme Inhibitors; Germination; Gibberellins; Hordeum; Okadaic Acid; Phosphorylation; Protoplasts; Seeds; Signal Transduction

Most of the inflammatory and proviral effects of tumor necrosis factor (TNF) are mediated through the activation of the nuclear transcription factor NF-kappa B. How TNF activates NF-kappa B, however, is not well understood. We examined the role of protein phosphatases in the TNF-dependent activation of NF-kappa B. Treatment of human myeloid ML-1a cells with TNF rapidly activated (within 30 min) NF-kappa B; this effect was abolished by treating cells with inhibitors of protein-tyrosine phosphatase (PTPase), including phenylarsine oxide (PAO), pervanadate, and diamide. The inhibition was dependent on the dose and occurred whether added before or at the same time as TNF. PAO also inhibited the activation even when added 15 min after the TNF treatment of cells. In contrast to inhibitors of PTPase, okadaic acid and calyculin A, which block serine-threonine phosphatase, had no effect. The effect of PTPase inhibitors was not due to the modulation of TNF receptors. Since both dithiothreitol and dimercaptopropanol reversed the inhibitory effect of PAO, critical sulfhydryl groups in the PTPase must be involved in NF-kappa B activation by TNF. PTPase inhibitors also blocked NF-kappa B activation induced by phorbol ester, ceramide, and interleukin-1 but not that activated by okadaic acid. The degradation of I kappa B protein, a critical step in NF-kappa B activation, was also abolished by the PTPase inhibitors as revealed by immunoblotting. Thus, overall, we demonstrate that PTPase is involved either directly or indirectly in the pathway leading to the activation of NF-kappa B.

Topics: Adenosine Triphosphate; Arsenicals; Base Sequence; Ceramides; Diamide; DNA-Binding Proteins; Dose-Response Relationship, Drug; Ethers, Cyclic; Humans; I-kappa B Proteins; In Vitro Techniques; Interleukin-1; Marine Toxins; Molecular Sequence Data; NF-kappa B; NF-KappaB Inhibitor alpha; Okadaic Acid; Oligodeoxyribonucleotides; Oxazoles; Protein Tyrosine Phosphatases; Signal Transduction; Tetradecanoylphorbol Acetate; Time Factors; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha; Vanadates

In response to insulin, several proteins are phosphorylated on tyrosine and on serine/threonine residues. Decreased phosphorylation of signaling peptides by a defective insulin receptor kinase may be a cause of insulin resistance. Accordingly, inhibition of the appropriate phosphatases might increase the phosphorylation state of these signaling peptides and thereby elicit increased glucose transport. The purpose of this study was to examine the effect of the serine/threonine phosphatase inhibitor okadaic acid and the tyrosine phosphatase inhibitors phenylarsine oxide and vanadate on 2-deoxyglucose transport in insulin-resistant human skeletal muscle. All three phosphatase inhibitors stimulated 2-deoxyglucose transport in insulin-resistant skeletal muscle. These data suggest that these compounds have bypassed a defect in at least one of the signaling pathways leading to glucose transport. Furthermore, maximal transport rates induced by the simultaneous presence of insulin and phosphatase inhibitor in insulin-resistant muscle were equal to insulin-stimulated rates in lean control subjects. However, both vanadate alone and vanadate plus insulin stimulated 2-deoxyglucose transport significantly more in insulin-sensitive tissue than in insulin-resistant tissue. These results demonstrate that although vanadate is able to stimulate glucose transport in insulin-resistant muscle, it is not able to normalize transport to the same rate achieved in insulin-sensitive muscle.

Topics: Adult; Animals; Arsenicals; Biological Transport; Deoxyglucose; Ethers, Cyclic; Humans; Insulin Resistance; Male; Muscle, Skeletal; Okadaic Acid; Phosphoprotein Phosphatases; Rats; Rats, Sprague-Dawley; Vanadates

Stimulation of phospholipase D after activation of cell surface receptors has been reported in many cell types. We have investigated the mechanism of activation of this enzyme by collagen in the human platelet by assaying the release of [3H]methylcholine from [3H]methylphosphatidylcholine. Results from these studies suggest that phospholipase D activity is regulated by reversible phosphorylation. Phospholipase D activity was stimulated when platelet-rich plasma was preincubated with collagen and was not inhibited by aspirin. Among various aggregating agents tested, collagen and thrombin but not ADP activated phospholipase D activity (2- to 3-fold). The addition of sphingosine inhibited phospholipase D activity. Preincubation of platelet-rich plasma with sphingosine inhibited collagen- and thrombin-induced platelet aggregation and the release of ATP. The inhibitory effect of sphingosine on collagen- and thrombin- induced platelet aggregation and release of ATP was dose-dependent. The functional significance of phospholipase D activation was also tested by examining the effect of the product, phosphatidic acid, on collagen-induced platelet aggregation and release of ATP. Platelet shape change and the reversibility of platelet aggregation resulted by the addition of phosphatidic acid to platelet-rich plasma. Furthermore, the simultaneous addition of phosphatidic acid and collagen shortened the latency period but had no effect on platelet aggregation. Two platelet proteins (47 kDa and 22 kDa) increased in phosphorylation after the addition of 1 microM phosphatidic acid which did not cause platelet aggregation. These results suggest that collagen stimulates phospholipase D activity which plays a secondary role in platelet aggregation and the release reaction.

Topics: Adenosine Triphosphate; Arsenicals; Blood Platelets; Collagen; Enzyme Activation; Ethers, Cyclic; Humans; Okadaic Acid; Phospholipase D; Platelet Aggregation; Platelet Aggregation Inhibitors; Sphingosine; Thrombin

Phenylarsine oxide (PAO), an inhibitor of tyrosine phosphatases, has been found to inhibit the early elevation in cytosolic Ca2+ concentration ([Ca2+]i), related to the CD3 activation pathway in Jurkat T cells. This inhibition was dose-dependent, consistent with previously reported effects of PAO on tyrosine phosphatases, and reversed by dimercaptopropanol. By contrast, okadaic acid, an inhibitor of serine/threonine phosphatases, had no effect on CD3-induced Ca2+ flux. PAO was compared with phorbol 12-myristate 13-acetate (PMA), which caused a similar, although less potent, inhibition as previously described. The two reagents produced additive inhibition of the CD3-induced [Ca2+]i rise, but did not affect thapsigargin- or ionomycin-driven Ca2+ flux in Jurkat cells. PAO and PMA prevented cells from complete depletion of intracellular Ca2+ stores by an anti-CD3 monoclonal antibody (mAb) and restored, at least partially, the ionomycin-sensitive pool, when added after anti-CD3 mAb. Moreover, the CD3-induced inhibition of phosphatidylserine synthesis, due to depletion of internal Ca2+ stores, is reversed by PAO and PMA. Anti-phosphotyrosine immunoblot analysis show that these effects cannot be accounted for by an inhibition of CD3-induced tyrosine phosphorylations. We propose that PAO and, to a lesser extent, PMA allow the refilling of internal compartments by Ca2+, which consequently abrogates a capacitative entry of external Ca2+.

Topics: Arsenicals; Biological Transport; Calcium; CD3 Complex; Cell Line; Cytosol; Ethers, Cyclic; Humans; Ionomycin; Okadaic Acid; Phosphatidylserines; Phosphorylation; T-Lymphocytes; Terpenes; Tetradecanoylphorbol Acetate; Thapsigargin; Tyrosine

The aim of this study was to investigate the stimulating effects of sulfhydryl reagents on glucose transport in isolated rat heart muscle cells and to compare them with the action of insulin. Low concentrations of the sulfhydryl oxidants hydrogen peroxide (H2O2) and diamide (5-100 microM), but also of phenylarsine oxide (PAO) (0.5-3 microM), that is known to specifically react with vicinal SH-groups, stimulated the rate of 2-deoxy-D-glucose uptake by a factor of 4 to 8 in these cells, while higher concentrations were inhibitory. The stimulating effects of H2O2 or diamide, and, to a significantly lesser extent, those of PAO or insulin, were depressed in cells pretreated with the sulfhydryl-alkylating agent N-ethylmaleimide (56-100 microM). H2O2 raised the Vmax and lowered the Km of 3-O-methyl-D-glucose uptake, while PAO or insulin solely increased Vmax. The increase in glucose transport caused by H2O2 was antagonized by the beta-adrenergic agonist isoprenaline (1 microM) or by a membrane-permeant cyclic AMP analog, whereas the effects of PAO or insulin were not altered. The action of H2O2 was additive with the stimulation induced by the protein phosphatase inhibitors okadaic acid (1 microM) or vanadate (6 mM), whereas the responses to PAO or insulin were reduced in the presence of these agents. Finally, H2O2 and PAO, but not insulin, acted additively with the protein kinase C ligand phorbol myristate acetate (0.8 microM) and with phospholipase C (0.03 units/ml). We conclude that, in cardiac myocytes, H2O2, on the one hand, and PAO (and possibly insulin), on the other hand, stimulate glucose transport via at least two distinct, SH-dependent pathways. These pathways, in turn, differ from a protein kinase C- and from a phospholipase C-mediated mechanism.

Topics: 3-O-Methylglucose; 8-Bromo Cyclic Adenosine Monophosphate; Animals; Arsenicals; Biological Transport; Cells, Cultured; Deoxyglucose; Dose-Response Relationship, Drug; Ethers, Cyclic; Ethylmaleimide; Glucose; Heart; Hydrogen Peroxide; Insulin; Methylglucosides; Myocardium; Okadaic Acid; Rats; Rats, Sprague-Dawley; Tetradecanoylphorbol Acetate; Vanadates

On a formal level the clocks regulating circadian and cell division cycles are related in that both have been modeled as limit cycle oscillations (Science, 211 (1981) 1002-1013; Brain Res., 504 (1989) 211-215; Proc. Natl. Acad. Sci. USA, 88 (1991) 7328-7332). Furthermore, in several organisms each clock system is able to modulate the other (Science, 211 (1981) 1002-1013). However, in spite of the similarities at the formal level, and the connections at the physiological level, no common cellular elements have been identified linking the two processes. In the current series of experiments we show that one key element of cell cycle regulation, tyrosine phosphorylation/dephosphorylation is intimately associated with circadian rhythm generation in the eye of the marine snail, Bulla gouldiana. The importance of tyrosine kinase activity in the generation of circadian rhythms provides a possible point of similarity between the fundamental biochemical mechanisms underlying both circadian and cell cycle clocks.

Topics: Animals; Arsenicals; Biological Clocks; Circadian Rhythm; Ethers, Cyclic; Eye; Genistein; Isoflavones; Okadaic Acid; Phosphates; Phosphoproteins; Phosphoric Monoester Hydrolases; Phosphorylation; Protein-Tyrosine Kinases; Snails; Tyrosine