The action of cyclic-AMP on GA3 controlled responses III. Characteristics of barley endosperm acid phosphatase induction by gibberellic acid and cyclic 3', 5'-adenosine monophosphate

Studies on the induction of barley endosperm acid phosphataseby GA₃ and cyclic-AMP revealed (1) acid phosphatase inductionby GA₃ and cyclic-AMP is inhibited in a similar manner by ABA,cycloheximide and 6-methyl purine, (2) combinations of GA₃ andphosphodiesterase inhibitors such as caffeine and theophyllinepromoted the level of acid phosphatase induction obtainablewith the hormone alone, and (3) cyclic-AMP and GA₃ may be actingas triggers in the induction of this enzyme. Some of the implicationsof these results are discussed. (Received May 25, 1972; )     

The action of cyclic-AMP on GA3 controlled responses II. Similarities in the induction of barley endosperm ATPase activity by gibberellic acid and cyclic 3',5'-adenosine monophosphate

The effect of cyclic-AMP and GA3 on ATPase production in embryolessbarley half-seeds was examined. Similarities in the responseinduced by 5 mM cyclic-AMP and GA₃ were observed in (1) thetotal amount of ATPase produced, (2) the time course for therelease of ATPase, and (3) the effect of abscisic acid on ATPaseinduction by these compounds. Of a variety of adenine nucleotidesexamined only cyclic-AMP was found to promote high levels ofATPase activity. Some of the implications of these results arediscussed. (Received May 11, 1971; )     

Cyclic 3', 5'-adenosine monophosphate phosphodiesterase mutants of Salmonella typhimurium.

Positive selection procedures for mutants of Salmonella typhimurium lacking cyclic 3', 5'7-adenosine monophosphate (cAMP) phosphodiesterase have been devised. The gene (cpd) coding for this enzyme has been located on the chromosome and shown to be 25% co-transducible with metC using phage P22. The mutants have been used to investigate the role of the enzyme in the control of genes whose expression is known to be dependent on cAMP. Significant alterations in the regulation of some but not others of these genes have been observed in these mutants. Mutants lacking the cAMP phosphodiesterase are more sensitive than their parents to a variety of antibiotics that appear to enter the cell through cAMP-dependent transport systems. They grow faster than the wild type on succinate-ammonia-salts, and glucose-proline-salts media and are inhibited by added cAMP on glucose, citrate, or glycerol-ammonia salts media whereas the wild type is unaffected. Neither the growth of Salmonella typhimurium on glycerol or citrate media nor the level of acid hexose phosphatase in the strain is affected by the loss of cAMP phosphodiesterase. In addition, the mutant strains are extremely sensitive to high levels of cAMP. Loss of the cAMP phosphodiesterase in strains unable to synthesize cAMP (adenyl cyclase negative) reduces by 10-fold the requirement for exogenous cAMP for expression of catabolite-sensitive phenotypes. These results suggest that through its control of cAMP levels in the cell the phosphodiesterase may be involved in the regulation of certain classes of catabolite-sensitive operaons and also in protecting the cell against high levels of cAMP.     

Effects of gibberellic acid and cyclic 3',5'-adenosine monophosphate on the flowering of Eschscholtzia californica Cham., a qualitative long day plant.

GA₃ and cyclic AMP induce floral buds in Eschscholtzia californicaunder noninductive short days and hasten it under inductivelong day conditions. (Received June 8, 1976; )     

The action of cyclic-AMP on GA3 controlled responses I. Induction of barley endosperm protease and acid phosphatase activity by cyclic-3',5'-adenosine monophosphate

The effect of cyclic-AMP on the induction of protease and acidphosphatase activity in embryoless barley half-seeds was examined.Promotion of protease synthesis was observed with 5 mu cyclic-AMPand ADP. The protease induced by these compounds was similarto protease obtained with GA₃, in that the enzyme was dependenton ß-mercaptoethanol for enhanced activity and increasedprotease activity observed in the media after a 48 hr incubationwith cyclic-AMP and ADP was due to an increase in both the synthesisand release of this enzyme. Of a variety of adenine compoundsand nucleoside diphosphates tested only cyclic-AMP and ADP inducedprotease activity. When these same compounds were tested fortheir ability to induce acid phosphatase activity only cyclic-AMPwas consistently found to be active. The total amount of enzymeproduced as well as the rate of enzyme release was similar forhalf-seeds incubated with 5 mu cyclic-AMP or a saturating GA3concentration. Inhibition of the hormone or nucleotide inducedpromotion of acid phosphatase was observed when abscisic acidwas added to the incubation medium with one of these compounds.Some of the implications of these results are discussed. (Received May 11, 1971; )     

Effect of cyclic 3',5'-adenosine monophosphate on central neurons

In experiments on unanesthetized non-immobilized rabbits, unit responses in the cortex and thalamus to the cyclic 3',5'-adenosine monophosphate (cAMPh) were studied by means of microionophoresis. It was shown that cAMPh changes the pattern of background unit activity, increasing or decreasing the discharge frequency. cAMPh changes unit responses both to acetylcholine and noradrenaline. These data permit, to assume that cAMPh participates both in adrenergic and in acetylocholinergic mechanisms of excitation processing in brain neurones.     

Stimulation of gibberellin-induced germination in lettuce seeds by cyclic 3',5'-adenosine monophosphate

Cyclic 3',5'-adenosine monophosphate and sodium dibutyryl cyclic3',5'-adenosine monophosphate had no effect on dark-germinationof light-sensitive lettuce {Lacluca sativa L., var. Grand Rapids)seeds when given alone. Cyclic 3',5'-adenosine monophosphate,however, synergistically enhanced the 3 µ 10^–5 Mgibberellic acid-induced germination, but showed no such effecton kinetin-stimulated germination. 5'-AMP, in contrast to cyclic3',5'-adenosine monophosphate, had no effect on gibberellicacidinduced germination. (Received August 8, 1971; )     

Measurement of cyclic 3',5'-adenosine monophosphate synthesis in growing Escherichia coli.

The net synthesis of cAMP by an adenine auxotroph of $\text{Escherichia coli}$ was measured by assaying the incorporation of tritium from [³H]-adenine into cyclic [³H] AMP during exponential growth. Synthesis of cAMP ceased abruptly when glucose was added to cells growing in glycerol and then recovered to an intermediate rate of synthesis after 0.5–1.0 generation. Cyclic AMP appeared to be synthesized from a precursor pool that turned over more rapidly than total cellular ATP. The rates of cAMP synthesis measured by this technique are compatible with the cellular levels of cAMP previously measured in this strain(3).     

The action of cyclic-AMP on GA3 controlled responses V. Similarities in the induction of isocitrate lyase in hazel seeds by gibberellic acid and cyclic 3',5'-adenosine monophosphate

Cyclic-AMP was found to promote the induction of isocitratelyase in hazel seeds. Similarities in the induction of thisenzyme by GA₃ and cyclic-AMP were found in the (1) total amountof enzyme produced, (2) the time course for the production ofthis enzyme, (3) the amount of enzyme secreted into the medium,and (4) the inhibition of enzyme production induced by ABA,cycloheximide and 6-methyl purine. Of a variety of nucleotidestested only cyclic-AMP consistently promoted high levels ofisocitrate lyase. Cyclic-AMP also promoted a similar increase in the fresh weightof the embryo to that obtained with GA₃ treated seeds. Thisinitial study is suggestive of the possibility that cyclic-AMPmay be involved in GA₃ mediated responses in hazel seeds. (Received May 4, 1973; )     

Cyclic 3',5'-adenosine monophosphate in the choroid plexus: stimulation by cholera toxin.

Cholera toxin was found to induce high accumulations of cyclic AMP in the isolated choroid plexus of the rabbit and in the incubation medium. The accumulation showed a characteristic lag phase of at least 30 min and continued for at least 3 hours. Inactivated cholera toxin was unable to increase cyclic AMP levels. There was only a moderate effect of cholera toxin on cyclic AMP “low K_m” phosphodiesterase activity in homogenates. The effect of cholera toxin on cyclic AMP levels confirms the existance of a potent cyclic AMP generating system in the choroid plexus which is activated also by β-adrenergic agonists, histamine and prostaglandin E₁.     

Cyclic 3',5'-adenosine monophosphate stimulates trehalose degradation in baker's yeast

The levels of cyclic 3′,5′-AMP and trehalose, as well as the specific activity of the trehalase have been investigated in cells of baker's yeast ($\text{Saccharomyces cerevisiae}$) during the lag phase preceding growth. During the first few minutes a substantial increase in the intracellular concentration of cyclic 3′,5′-AMP was observed, followed by a 6–8 fold increase in trehalase activity concomitant with the rapid degradation of trehalose. Cell free extracts prepared from resting yeast were shown to contain a cryptic trehalase, which under physiological conditions could be activated by cyclic 3′,5′-AMP to the same degree as $\text{in vivo}$. These observations suggest that in the lag phase of growth, the level of trehalose in baker's yeast is under control of a system, regulated by the level of cyclic 3′,5′-AMP.     

Promotion and inhibition of {alpha}-amylase production in barley endosperm by cyclic 3',5'-adenosine monophosphate and adenosine diphosphate

The possibility that gibberellin-induced -amylase synthesisin barley endosperm might be mediated by cyclic-3',5'-adenosinemonophosphate (3',5'-AMP) was examined. Promotion of -amylasesynthesis by 3',5'-AMP (5 mM) was observed in the absence ofgibberellic acid (GA₃) and in combination with GA₃ at concentrationsbelow 2 mµM. When combined with gibberellin at concentrationsabove 2 mµM, however, 3',5'-AMP reduced the amount of-amylase obtained. The cyclic nucleotide showed slight activityat concentrations as low as 0.05 mM. These promotions were shownto be due to increased synthesis of -amylase rather than toan increased secretion of the enzyme. Of a variety of adeninecompounds and nucleoside diphosphates tested only 3',5'-AMPand adenosine diphosphate (ADP) induced -amylase synthesis.Longer incubation times were necessary to obtain maximal -amylaseinduction with the nucleotides than with GA₃. ADP and 3',5'-AMPwere about one third and one fifth as active, respectively,as GA₃ in promoting -amylase synthesis, although GA₃ was morethan 10⁷ times more effective. AMO-1618 did not inhibit theaction of the nucleotides and methanolic extracts of the nucleotidesshowed no gibberellin-like activity. Both nucleotides were synergisticwith GA₃ in overcoming the inhibitory effects of acetate andcitrate buffers on -amylase synthesis. (Received February 24, 1969; )