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).     

Regulation of beta-galactosidase synthesis in Escherichia coli by exogenous cyclic 3',5'-adenosine monophosphate]

The effect of cyclic 3',5'-adenosine monophosphate (cAMP) on the rate of beta-galactosidase biosynthesis was studied in the cells of Escherichia coli M-17 growing in MPB and mineral media with glucose and maltose, i.e. under the conditions of various catabolite repression, as well as upon lac-operon induction by isopropyl-beta-D-galactopyranoside (IPGP). The stimulating action of exogenous cAMP was found only in a medium with salts and glucose. The induction by IPGP was highest during the growth in a medium with glucose and maltose. When the medium contained IPGP, cAMP accelerated the enzyme synthesis in all media, but only at the early growth phases, while cAMP eliminated the effect of IPGP at the stationary phase of growth. The regulation of beta-galactosidase biosynthesis by cAMP demonstrated for the first time that this effect depended on the physiological state of E. coli: the expression of catabolite-sensitive E. coli genes was subject to both positive and negative regulation in one and the same inducible system. The effect exerted by cAMP depended on the nature of a carbon source in the growth medium.     

Effect of cyclic 3',5'-adenosine monophosphate on the growth rate of Escherichia coli

Cyclic 3',5'-adenosine monophosphate (cAMP) inhibits the rate of Escherichia coli growth in media with glucose. When the exogenous nucleotide is added, the generation time and the lag phase become longer. These parameters decrease if cAMP is entirely absent from the cya- mutant as compared to the parent cya+ strain. The nucleotide exerts a low activity in media with glycerol. The action of cAMP is highly specific.     

Influence of cyclic 3',5'-adenosine monophosphate on uracil uptake by rifampicin treated Escherichia coli cells.

Incubation of cells from a wild type strain of E. coli with 0.3 mg/ml rifampicin for 15 minutes lead to a complete inhibition of RNA synthesis measured as the uracil incorporation into the trichloroacetic acid insoluble fraction. In these rifampicin-treated cells [14C]uracil incorporation tended to decrease during a further incubation at 37 degrees. Addition of cyclic AMP increased the inactivation of the system responsible for [14C]uracil uptake. The cyclic nucleotide effect seems to be specific since ATP or 5'AMP did not increase such inactivation.     

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; )     

Effect of cyclic 3',5'-adenosine monophosphate on the sporulation of Saccharomyces cerevisiae

Cyclic 3',5'-adenosine monophosphate and sodium dibutyryl cyclic3',5'-adenosine monophosphate had no effect on sporulation ofSaccharomyces cerevisiae, when added to a sporulation mediumnot enriched with glucose. They did, however, reverse the repressionof sporulation by glucose, when added to the sporulation mediumtogether with glucose. 5'-AMP, 5'-ADP and 5'-ATP did not reversethe repression of sporulation by glucose. (Received February 24, 1972; )     

Stimulation of galactokinase synthesis in Escherichia coli by adenosine 3',5'-cyclic monophosphate

Adenosine 3',5'-cyclic monophosphate (cyAMP) stimulates the rate of synthesis of galactokinase in glycerol-grown Escherichia coli both when production of the enzyme is induced by d-fucose and when it is repressed by glucose in the presence of inducer. cyAMP also stimulates the synthesis of galactokinase in constitutive strains B78A (R(-)) and R10 (O(c)), and overcomes the transient repression of galactokinase synthesis caused by glucose.     

The cyclic 3'',5''-adenosine monophosphate receptor protein and regulation of cyclic 3'',5''-adenosine monophosphate synthesis in Escherichia coli.

Summary Rates of synthesis of cyclic 3,5-adenosine monophosphate (cAMP) were measured in cultures of Escherichia coli aerating without a carbon source. This technique provides a representative measure of adenylate cyclase activity in the absence of inhibition caused by transport of the carbon source. Adenylate cyclase activity was found to vary more than 20-fold depending on the carbon source that had been available during growth. Synthesis of cAMP in cells aerating in the absence of the carbon source was highest when cells had been grown with glucose or fructose which inhibit adenylate cyclase activity severely. Synthesis of cAMP was much lower when cells had been grown with glycerol or succinate which cause only minimal inhibition of the activity.The variation in cAMP synthesis due to different carbon sources requires a functional cAMP receptor protein (CRP). Crp^- mutants synthesize cAMP at comparable rates regardless of the carbon source that afforded growth. A novel mutant of E. coli having a CRP no longer dependent on cAMP has been isolated and characterized. Adenylate cyclase activity in this mutant no longer responds normally to variations in the carbon source.