Catalytic Activity Of Adenylate Cyclase Research Articles

Behavior of Hypertrophied Right Ventricle during the Development of Left Ventricular Failure Due to Myocardial Infarction.

In order to determine the behavior of the right ventricle, we have reviewed the existing literature in the area of cardiac remodeling, signal transduction pathways, subcellular mechanisms, β-adrenoreceptor-adenylyl cyclase system and myocardial catecholamine content during the development of left ventricular failure due to myocardial infarction. The right ventricle exhibited adaptive cardiac hypertrophy due to increases in different signal transduction pathways involving the activation of protein kinase C, phospholipase C and protein kinase A systems by elevated levels of vasoactive hormones such as catecholamines and angiotensin II in the circulation at early and moderate stages of heart failure. An increase in the sarcoplasmic reticulum Ca2+ transport without any changes in myofibrillar Ca2+-stimulated ATPase was observed in the right ventricle at early and moderate stages of heart failure. On the other hand, the right ventricle showed maladaptive cardiac hypertrophy at the severe stages of heart failure due to myocardial infarction. The upregulation and downregulation of β-adrenoreceptor-mediated signal transduction pathways were observed in the right ventricle at moderate and late stages of heart failure, respectively. The catalytic activity of adenylate cyclase, as well as the regulation of this enzyme by Gs proteins, were seen to be augmented in the hypertrophied right ventricle at early, moderate and severe stages of heart failure. Furthermore, catecholamine stores and catecholamine uptake in the right ventricle were also affected as a consequence of changes in the sympathetic nervous system at different stages of heart failure. It is suggested that the hypertrophied right ventricle may serve as a compensatory mechanism to the left ventricle during the development of early and moderate stages of heart failure.

In vivo induction of CTL responses by recombinant adenylate cyclase of Bordetella pertussis carrying viral CD8+ T cell epitopes.

Exogenous Ags enter the endosomal pathway and are presented to CD4+ T cells in association with class II molecules whereas endogenously synthesized Ags, such as viral proteins, are presented to CD8+ T cells in association with MHC class I molecules. Therefore, most CTL activation strategies use live vectors although an alternative possibility could be to deliver the epitope into the cytosol by targeting it to an invasive nonreplicative vector. The adenylate cyclase toxin of Bordetella pertussis is able to invade a large number of eukaryotic cells and to deliver its catalytic domain to the cytosol of the cells. In the present study, we have tested the in vivo immunogenicity of recombinant adenylate cyclase toxins expressing CTL epitopes either from the nucleoprotein of lymphocytic choriomeningitis virus or from the V3 region of HIV-1 gp120. BALB/c mice immunized with these toxins developed high specific CTL responses that were shown to be mediated by class I-restricted CD8+ CTL. The induction of CTL responses by recombinant toxins did not require CD4+ T cells and the cytotoxic activity persisted 2 mo after immunization. The activation of CTL responses by the recombinant adenylate cyclase toxin required the full-length invasive activity of the toxin but did not depend upon its catalytic adenylate cyclase activity as demonstrated with a genetically inactivated recombinant toxin expressing the lymphocytic choriomeningitis virus epitope. This genetically detoxified invasive toxin represents, therefore, an attractive new vector for CTL activation.

Effect of diabetes mellitus on responses of the rat basilar artery to activation of beta-adrenergic receptors

The goal of this study was to determine whether diabetes mellitus alters reactivity of the basilar artery in vivo to activation of beta-adrenergic receptors. We measured diameter of the basilar artery in non-diabetic and diabetic (streptozotocin 50–60 mg/kg i.p.) rats during superfusion with isoproterenol and norepinephrine, which dilate cerebral blood vessels via activation of beta-adrenergic receptors. Dilatation of the basilar artery in response to isoproterenol and norepinephrine was significantly less in diabetic compared to non-diabetic rats. To determine whether dmpaired dilatation of the basilar artery in diabetic rats in response to isoproterenol and norepinephrine was related to an alteration in catalytic activity of adenylate cyclase, we examined dilatation of the basilar artery in response to forskolin, a direct activator of adenylate cyclase. In contrast to that observed for isoproterenol and norepinephrine, forskolin produced similar dilatation of the basilar artery in non-diabetic and diabetic rats. Thus, the findings of the present study suggest that diabetes mellitus impairs dialtation of the basilar artery in vivo in response to activation of beta-adrenergic receptors. In addittion, impairment of beta-adrenergic mediated dilatation of the basilar artery during diabetes mellitus does not appear to be related to an alteration in catalytic activity of adenylate cyclase.

Intraventricular changes in the beta-adrenoceptor-adenylate cyclase system of the rat heart with the progress of monocrotaline-induced right ventricular hypertrophy.

We investigated the changes in the membranous beta-adrenoceptor-adenylate cyclase system in the right ventricle, left ventricle and interventricular septum during the progress of monocrotaline-induced right ventricular hypertrophy and failure. The beta-adrenoceptor density was decreased in hypertrophied right ventricle 2 to 4 weeks after treatment. When the rats showed symptoms of right ventricular failure 4 weeks after treatment, the beta-adrenoceptor density was decreased in the interventricular septum. Both basal and forskolin-stimulated adenylate cyclase activities were decreased in the right ventricle at 3 and 4 weeks, and in the interventricular septum at 4 weeks, after treatment, which indicates that the catalytic activity of adenylate cyclase is reduced. Changes in isoproterenol plus Gpp (NH) p- or sodium fluoride-stimulated adenylate cyclase activity were generally similar to those in basal activity. These data indicate that a chamber-specific decrease in beta-adrenoceptor density begins in the early stages of right ventricular hypertrophy, and that beta-adrenoceptor density and adenylate cyclase activity in the interventricular septum are decreased in the advanced stages of heart failure in monocrotaline-treated rats.

Regional mapping of a human rodα-transducin (GNAT1) gene to chromosome 3p22

Guanine nucleotide binding proteins (G-proteins) function as second messengers to modulate several cellular processes in response to external stimuli. The most extensively studied G-proteins are GNAT1, GNAS1, and GNAI1. The gene product of GNAT1 activates cGMP phosphodiesterase by coupling to photolyzed rhodopsin, while gene products of GNAS1 or GNAI1 stimulate or inhibit adenylate cyclase catalytic activity in hormone-sensitive systems, respectively. All three proteins are heterotrimers composed of alpha, beta, and gamma subunits, and the individual subunits share similar enzymatic properties. The alpha subunits bind guanyl nucleotides and catalyze the hydrolysis of GTP to GDP and Pi. In addition to GNAT1, which has been localized to chromosome 3, other alpha G-proteins have also been cloned and mapped. Because of its exclusive retinal expression, regional mapping of the GNAT1 locus would facilitate linkage studies of inherited retinal diseases. Using in situ hybridization, the authors have localized the GNAT1 locus to 3p22.

Modulation by sex hormones of the membranous transducing system regulating fatty acid mobilization in adipose tissue

This review summarizes recent animal studies performed to determine the possible role played by sex hormones in the sex- and site-related differences characterizing adipocyte lipolytic activity. In both normal female rats and male hamsters, fat cells from deep intra-abdominal sites elicit higher catecholamine-stimulated lipolytic responses than subcutaneous adipocytes. By using ovariectomized rats, it was found that estradiol ‘in vivo’, while having no effect in subcutaneous cells, promotes catecholamine-stimulated lipolysis in deep intra-abdominal adipocytes by increasing their adenylate cyclase catalytic activity. By using castrated hamsters, it was found that both deep intra-abdominal and subcutaneous fat cell lipolytic activities are equally sensitive to testosterone. In these cells, testosterone treatment promotes not only the β-adrenergic lipolytic responses by increasing both the adenylate cyclase catalytic activity and the Gsα level, but also enhances the α2-adrenergic antilipolytic responses through a transcriptional activation of the α2-adrenoceptor expression. These experiments demonstrate that in some, but not all, white adipocytes the adrenergic signal transducing system regulating lipolysis is a target for sex hormones. This finding may have potential importance in the understanding of the mechanisms underlying the sex-related regional specificities of adipose tissue metabolism and distribution.

Targeted mutations that ablate either the adenylate cyclase or hemolysin function of the bifunctional cyaA toxin of Bordetella pertussis abolish virulence.

Bordetella pertussis, the causative agent of whooping cough, secretes several toxins implicated in this disease. One of these putative virulence factors is the adenylate cyclase (AC) toxin that elevates intracellular cAMP in eukaryotic cells to cytotoxic levels. This toxin is a bifunctional protein comprising both AC and hemolysin (HLY) enzymatic domains. The gene encoding the AC toxin (cyaA) is expressed as part of an operon that includes genes required for secretion or activation of the toxin. Because of this genetic organization, it is difficult to create B. pertussis mutants of cyaA that are ablations of a single enzyme function by conventional means, such as transposon mutagenesis. Therefore, to clarify the role of individual toxin functions in the virulence of B. pertussis, we have used site-directed or deletion mutagenesis and genetic recombination to specifically target the cyaA gene of B. pertussis to produce mutants that lack only the AC or HLY activity of this toxin. A point mutant of B. pertussis with abolished AC catalytic activity was greater than 1000 times less pathogenic to newborn mice than wild-type bacteria, directly demonstrating the importance of the AC toxin in pertussis virulence. Similarly, an in-frame deletion mutant of B. pertussis that lacks HLY is equally avirulent, supporting observations that the HLY domain plays a critical role in AC toxin entry into cells. Furthermore, the genetically inactivated AC toxin produced by the point mutant is antigenically similar to the native toxin, suggesting that this strain may be useful in the development of pertussis component vaccines.

Differential modulation of the adenylate cyclase/cyclic AMP stimulatory pathway by protein kinase C activation in rat adipose tissue and isolated fat cells: Influence of collagenase digestion

Exposure of rat epididymal fat pad to phorbol 12-myristate 13-acetate (TPA), an activator of protein kinase C, results in an 85% increase in isoproterenol-stimulated cyclic AMP (cAMP) accumulation, an effect which was antagonized by H7, a protein kinase C inhibitor. This promoting action of TPA appears to be related to (i) an increase in the catalytic activity of adenylate cyclase, (ii) an increase in the maximal response of adenylate cyclase to fluoride and guanylimidodiphosphate (GppNHp) with no change in the ec 50 value for GppNHp, and (iii) a reduction of the isoproterenol-stimulated low- K m cAMP phosphodiesterase activity present in the 30,000 g pellet of fat pad homogenates. In contrast with fat pads, exposure of isolated rat fat cells to TPA failed to influence their adenylate cyclase response to GppNHp and their cAMP accumulation and lipolysis. However, the other alterations caused by TPA in fat pads were still observed in fat cells. These results suggest that (i) the major alteration responsible for the promoted isoproterenol-stimulated cAMP response observed in fat pads after exposure to TPA is an increased interaction between the αs subunit of Gs and the catalytic site of adenylate cyclase and (ii) this increased interaction is dependent on protein kinase C activation and is abolished by collagenase digestion.

Dietary (n-6) Polyunsaturated Fatty Acids Affect β-Adrenergic Receptor Binding and Adenylate Cyclase Activity in Pig Adipocyte Plasma Membrane

The effect of dietary (n-6) polyunsaturated fatty acids (sunflower oil) on pig adipocyte beta-adrenoreceptor and adenylate cyclase activity was examined. Two adipose sites (subcutaneous and perirenal) were compared. The existence of two affinity classes for beta-adrenoreceptors was evidenced. Adenylate cyclase stimulation by isoproterenol was higher in the perirenal fat than in the subcutaneous fat, in parallel to a higher beta-adrenoreceptor density. When sunflower oil was included in the diet, the adenylate cyclase response to beta-agonists was greater, particularly in perirenal fat, as was the affinity of the adrenoreceptors in their high affinity state. However, the number of beta-adrenoreceptors was lower, suggesting that these are spare receptors. Adenylate cyclase stimulation by 5'-guanylylimidophosphate also revealed site- and diet-specific variations indicating alterations at the Gs-protein level. The adenylate cyclase catalytic activity (reflected by the forskolin-stimulated response) in the control group was higher in the subcutaneous fat than in the perirenal fat. In the sunflower oil-fed pigs, the catalytic activity was greater in the perirenal fat relative to controls, leading to similar values in both adipose tissues of sunflower oil-fed pigs. This indicates that the cyclase catalytic subunit activity also depends on the anatomical site of the fat deposit and is influenced by the diet as well. Correlation between these changes in the adenylate cyclase system are discussed in relationship with alterations in the plasma membrane structure.

Influence of Ovariectomy and Regional Fat Distribution on the Membranous Transducing System Controlling Lipolysis in Rat Fat Cells*

The mechanism by which sex steroid hormones modulate the lipolytic process in fat cells is still not completely understood, particularly at the level of the membrane. As estrogens seem to have regional influence on adipose tissue, we have compared the effects of long term ovariectomy (3 weeks) on the membranous transducing system controlling lipolysis in two rat fat deposits, the femoral sc and the parametrial adipose tissue. These experiments demonstrate the following. 1) Compared to parametrial adipocytes, sc adipocytes have a reduced cAMP production and a decreased beta-adrenoceptor number associated with a reduced lipolytic activity. 2) In parametrial adipocytes, ovariectomy induces a decrease in cAMP production and in both the beta-agonist-stimulated response and the catalytic activity of adenylate cyclase in association with a decreased lipolysis. In contrast, these parameters are unaltered by ovariectomy in sc adipocytes. This study, which reveals site-related differences in the sensitivity of the fat cell adenylate cyclase system to ovarian status, suggests that these differences may explain some of the sex-related regional specificities of adipose tissue metabolism and distribution.

Mechanosensitive adenylate cyclase activity in coronary vascular smooth muscle cells

The purpose of the present study was to test the hypothesis that adenylate cyclase activity of porcine coronary artery smooth muscle cells is sensitive to mechanical stretch. Cultured vascular smooth muscle cells were stretched at 24% maximal strain at 60 cycles/min for 30 minutes. Both basal and maximal catalytic activity of adenylate cyclase (as assessed by stimulation by 100 μM forskolin with 5 mM manganese chloride) were reduced by 30% (P<0.05) in membranes obtained from stretch versus unstretched cells. The magnitude of the stretch-induced reduction in Gpp(NH)p was identical over the entire time course studied (5–30 minutes). Furthermore, basal adenylate cyclase activity was inversely related to the magnitude of stretch. Thus, cyclic stretch can influence adenylate cyclase activity in coronary vascular smooth muscle cells. These data provide important information concerning potential biochemical mechanisms involved in the myogenic response of vascular smooth muscle and also suggest a potential mechanism by which the coronary circulation may adapt to chronically reduced perfusion pressure.

Correction by Dexamethasone Treatment of the Altered Lipolytic Cascade Induced by Adrenalectomy in Rat Adipocytes

The ability of glucocorticoid-treatment to reverse the metabolic alterations caused by adrenalectomy in rat adipocytes was studied. Correction of the enhanced adenosine antilipolytic effect and of the defect in lipolysis, cyclic AMP and adenylate cyclase responsiveness to guanine nucleotides were all achieved after a 24 h dexamethasone treatment, whereas correction of the defect in beta-adrenoceptor-density required a 48 h treatment. The latter treatment, however, failed to reverse the defect in both the adenylate cyclase catalytic activity and protein content per fat cell. These different kinetics of restoration indicate that correction by dexamethasone of the defective cyclic AMP and lipolytic responses on one hand and of the guanine nucleotide control of adenylate cyclase on the other one are two related phenomenoms.

Guanine nucleotide binding regulatory proteins and adenylate cyclase in livers of streptozotocin- and BB/Wor-diabetic rats. Immunodetection of Gs and Gi with antisera prepared against synthetic peptides.

Adenylate cyclase in liver plasma membranes from streptozotocin-diabetic (STZ) or BB/Wor spontaneously diabetic rats showed increased responsiveness to GTP, glucagon, fluoroaluminate, and cholera toxin. Basal or forskolin-stimulated activity was unchanged in STZ rats, but increased in BB/Wor rats. No change in the alpha-subunit of Gi (alpha i) was observed in STZ or BB/Wor rats using pertussis toxin-stimulated [32P]ADP-ribosylation. Immunodetection using antibodies against the COOH-terminal decapeptides of alpha T and alpha i-3 showed no change in alpha i in STZ rats and a slight decrease in BB/Wor rats. Angiotensin II inhibition of hepatic adenylate cyclase was not altered in either diabetic rat. In both models of diabetes, Gs alpha-subunits were increased as measured by cholera toxin-stimulated [32P]-ADP-ribosylation of 43-47.5-kD peptides, reconstitution with membranes from S49 cyc- cells or immunoreactivity using antibodies against the COOH-terminal decapeptide of alpha s. These data indicate that STZ-diabetes increases hepatic Gs but does not change Gi or adenylate cyclase catalytic activity. In contrast, BB/Wor rats show increased hepatic Gs and adenylate cyclase. These changes could explain the increase in hepatic cAMP and related dysfunctions observed in diabetes.

Progesterone depression of norepinephrine-stimulated cAMP accumulation in hypothalamic slices

The present experiments examined the effects of progesterone on adrenergic receptor coupling to adenylate cyclase in hypothalamic and preoptic area slices by monitoring norepinephrine (NE)-stimulated increases in cAMP accumulation. Progesterone treatment of estrogen-primed rats decreased NE-induced slice cAMP accumulation. The reduced cAMP response was estrogen-dependent since it was not demonstrable in slices from rats exposed to progesterone without prior estrogen priming. Neither generalized increases in phosphodiesterase activity nor decreases in the catalytic activity of adenylate cyclase could account for the reduced ability of NE to stimulate cAMP accumulation in hypothalamic slices. Moreover, the cAMP response to two other activators of adenylate cyclase, adenosine and vasoactive intestinal peptide, was not decreased in slices from rats treated with estrogen plus progesterone. Selective adrenergic agonists and antagonists were employed to determine which adrenergic receptors mediate cAMP accumulation in progesterone-exposed slices. Slice cAMP levels were elevated by the β receptor agonist isoproterenol but not by α 1 (phenylephrine) or α 2 (clonidine) agonists. However, clonidine potentiated the effect of isoproterenol on slice cAMP formation whereas phenylephrine did not. Likewise, NE-stimulated cAMP accumulation was completely antagonized only by a combination of both β (propranolol) and α 2 (yohimbine) antagonists. The data suggest that in slices from estrogen plus progesterone-treated rats, α 2 receptors contribute significantly to NE stimulation of cAMP accumulation. The overall depression of the cAMP response to NE in progesterone-exposed slices may involve a decrease of α 1 receptor facilitation of cAMP synthesis.

Role of Adenosine 3',5'-Monophosphate and the Ri-Receptor Gi-Coupled Adenylate Cyclase Inhibitory Pathway in the Mechanism Whereby Adrenalectomy Increases the Adenosine Antilipolytic Effect in Rat Fat Cells*

The aim of this study was to establish the mechanism by which adrenalectomy promotes the antilipolytic effect of the adenosine analog (-)-N6-(R-phenyl-isopropyl)adenosine (R-PIA) in rat fat cells. This action of adrenalectomy was not specific for R-PIA, since it was also observed with nicotinic acid and was prevented by phosphodiesterase inhibitors. In contrast, the inhibitory effect of R-PIA and nicotinic acid toward isoproterenol-stimulated cAMP accumulation was unaltered by adrenalectomy regardless of whether phosphodiesterase inhibitors were present. Whatever the conditions used, however, the cAMP levels in adrenalectomized rat adipocytes were one quarter to one third of those in sham-operated rats and remained below the limit over which variations in cAMP had no more influence in lipolysis. Both total and particulate low Km cAMP phosphodiesterase activities per adipocyte were decreased in adrenalectomized rats, but the stimulatory responses of the particulate enzyme to R-PIA remained unchanged. Pertussis toxin-catalyzed ADP ribosylation studies revealed a marked decrease in the total amount of the alpha-subunits of Go and the adenylate cyclase inhibitory regulatory protein Gi after adrenalectomy. However, the inhibitory dose-response curves of adenylate cyclase to R-PIA, nicotinic acid, GTP, guanylylimidodiphosphate, and guanosine 5'-O-(3-thiotriphosphate) were unaltered by adrenalectomy, indicating that the inhibitory function of Gi is unimpaired by adrenalectomy. Lastly, adrenalectomy resulted in a 60% reduction of the Mn2+-stimulated adenylate cyclase activity/adipocyte, which indicates that adrenalectomy causes a defect in adenylate cyclase catalytic activity. Thus, enhanced antilipolytic effects of R-PIA induced by adrenalectomy do not involve increased function of the adenosine receptor Gi-coupled adenylate cyclase inhibitory pathway, but are related to abnormally low intracellular cAMP levels due to defective adenylate cyclase catalytic activity.

Endothelium-mediated coronary dilatation by adenosine does not depend on endothelial adenylate cyclase activation: studies in isolated guinea pig hearts

Adenosine, applied to the coronary system of guinea pigs at up to 10(-6) M, elicits dilatation solely via an endothelium-mediated process. We investigated the role of coronary A2 receptors in this dilation, since the coronary endothelium possesses adenosine A2-receptors with a stimulatory action on the adenylate cyclase. In situ, A2 receptor stimulation can be assessed by prelabeling the coronary endothelial adenine nucleotide pool with 3H-adenosine and subsequently determining the rate of release of radiolabeled cAMP induced by A2 agonists. Thus, perfusion of isolated hearts with 5'-N-ethylcarboxamidoadenosine (NECA) dose-dependently increased coronary flow and the release of 3H-cAMP from the endothelium. In the presence of 50 microM 2',5'-dideoxyadenosine (ddA), a P-site agonist which inhibits the catalytic activity of adenylate cyclase, coronary flow increases induced by both adenosine and NECA were unaffected. In contrast, ddA reduced the release of labeled cAMP in response to NECA by about 60%. In cultured endothelial cells, ddA likewise inhibited cAMP accumulation due to NECA by about 70%. Moreover, ddA antagonized the adenylate cyclase mediated flow response due to the PGI2 analogue, iloprost, as well as the positive chronotropic and inotropic actions of isoproterenol. The dissociation elicited by ddA between the coronary flow response and the release of cAMP strongly indicates that the endothelial A2 receptors which are linked to adenylate cyclase are not causally involved in endothelium-dependent coronary dilatation induced by adenosine.

Increased adenylate cyclase catalytic activity explains how estrogens “in vivo” promote lipolytic activity in rat white fat cells

Estradiol administration (5 μg per day × 4 days) to ovariectomized rats resulted in a 60–70 % increase in the maximal lipolytic response of their white adipocytes to isoproterenol, epinephrine, IBMX and forskolin. These altered lipolytic responses were accompanied by parallel changes in the intracellular cyclic AMP levels found in response to 1 mM IBMX alone (+ 106 %) or combined with submaximal concentrations of isoproterenol (+ 205 %), epinephrine (+ 190 %) and forskolin (235 %). Studies of the adenylate cyclase activity revealed an overall increase in the stimulatory responsiveness of the enzyme (+ 150 to + 200 %) after the estradiol-treatment, regardless of the stimulatory agents tested (GTP, GppNHp, fluoride, isoproterenol, ACTH, forskolin). Finally, the finding of a 2-fold enhancement of the Mn 2+ (± GDP β S)-stimulated adenylate cyclase activity after the estradiol-treatment strongly suggests that increased activity of the catalytic subunit of this enzyme is the likely mechanism whereby estrogens promote lipolysis in rat fat cells.

Permissive action of glucocorticoids on catecholamine-induced lipolysis: Direct “in vitro” effects on the fat cell β-adrenoreceptor-coupled-adenylate cyclase system

Exposure of adipose tissue fragments to dexamethasone leads to enhanced lipolytic and cyclic AMP responses of isolated fat cells to isoproterenol. This permissive effect of the steroid is dose-dependent, prevented by the glucocorticoid receptor antagonist RU 38486, maximum after 48 h exposure to 10 nM dexamethasone and affects only the amplitude of the maximal response (+50%). Exposure to dexamethasone also induces an increase in both the number of beta-adrenergic receptors (+30%), and the adenylate cyclase-catalytic activity (+64%) and - responses to GTP (+114%) and isoproterenol (+55%). These data strongly suggest that the permissive effect of glucocorticoids towards lipolysis "in vivo" results at least in part from a glucocorticoid-receptor mediated action of these hormones on the fat cell membranous components involved in the beta-adrenergic control of lipolysis.

Restoration of Adenylate Cyclase Responsiveness in Murine Myeloid Leukemia Permits Inhibition of Proliferation by Hormone. Butyrate Augments Catalytic Activity of Adenylate Cyclase

Mechanisms of leukemic cell clonal dominance may include aberrations of transmembrane signaling. In particular, neoplastic transformation has been associated with reduced capacity for hormone-stimulated adenylate cyclase activity. In the present study, prostaglandin E, a hormonal activator of adenylate cyclase that has antiproliferative activity in myeloid cells, and cholera toxin, an adenylate cyclase agonist that functions at a postreceptor site by activating the adenylate cyclase stimulatory GTP-binding protein (Gs), were studied for antiproliferative activity in two murine myeloid cell lines. FDC-P1, an interleukin 3 (IL 3)-dependent myeloid cell line and a tumorigenic IL 3-independent subline, FI, were resistant to these antiproliferative agents. The in vitro ability of the "differentiation" agent, sodium butyrate, to reverse their resistance to adenylate cyclase agonists was studied. The antiproliferative action of butyrate involved augmentation of transmembrane adenylate cyclase activity. Increased adenylate cyclase catalyst activity was the primary alteration of this transmembrane signaling group leading to the functional inhibitory effects on leukemia cells, although alterations in regulatory G-proteins appear to play a secondary role.

Restoration of adenylate cyclase responsiveness in murine myeloid leukemia permits inhibition of proliferation by hormone. Butyrate augments catalytic activity of adenylate cyclase

Mechanisms of leukemic cell clonal dominance may include aberrations of transmembrane signaling. In particular, neoplastic transformation has been associated with reduced capacity for hormone-stimulated adenylate cyclase activity. In the present study, prostaglandin E, a hormonal activator of adenylate cyclase that has antiproliferative activity in myeloid cells, and cholera toxin, an adenylate cyclase agonist that functions at a postreceptor site by activating the adenylate cyclase stimulatory GTP-binding protein (Gs), were studied for antiproliferative activity in two murine myeloid cell lines. FDC-P1, an interleukin 3 (IL 3)-dependent myeloid cell line and a tumorigenic IL 3- independent subline, FI, were resistant to these antiproliferative agents. The in vitro ability of the “differentiation” agent, sodium butyrate, to reverse their resistance to adenylate cyclase agonists was studied. The antiproliferative action of butyrate involved augmentation of transmembrane adenylate cyclase activity. Increased adenylate cyclase catalyst activity was the primary alteration of this transmembrane signaling group leading to the functional inhibitory effects on leukemia cells, although alterations in regulatory G- proteins appear to play a secondary role.