SUMMARY: Streptomyces antibioticus produces a cell-wall-located deoxyriboendonuclease (DNAase) the synthesis of which in submerged and surface cultures is related to the growth rate. DNAase synthesis always preceded aerial mycelium formation in surface cultures. Production of aerial mycelium began at the end of exponential growth or in the early stationary phase; it was absent in cultures grown on nutrient agar/glucose or in media with a high concentration of casein hydrolysate. These nutritional conditions also impaired production of the DNAase. External DNA substrates were not degraded by mycelium producing the DNAase. These observations lead us to suggest a role for the enzyme in the developmental cycle of S. antibioticus.
SUMMARY: General properties of S-adenosylmethionine decarboxylase (SAMDC) from Mucor rouxii were studied. Dormant spores of the fungus did not contain detectable levels of the enzyme, but it started to be synthesized at early stages of spore germination. Kinetics of synthesis changed before emergence of the germ tube, with a corresponding increase in a second SAMDC activity which, in contrast to the one originally synthesized, was not activated by putrescine. Development of the second enzyme activity required de novo protein synthesis. Neither enzymic activity was stimulated by Mg2+. Addition of the SAMDC inhibitor methylglyoxal bis-(guanylhydrazone) (MGBG) stopped fungal development in growth phase Ia: cells became spherical and showed ultrastructural alterations. Although MGBG inhibited polyamine formation, it barely inhibited protein and RNA biosynthesis during the first hour of incubation. However, at later periods, biosynthesis of both macromolecules was strongly decreased. When MGBG was added to growth media 3 h after inoculation of spores, it did not affect spore germination and outgrowth. A hypothesis for two different roles of spermidine and putrescine in spore germination is discussed.
SUMMARY: Aspergillus nidulans produces an extracellular β-D-fructofuranoside fructohydrolase (invertase) when grown on a medium containing the β-fructofuranosides sucrose or raffinose, indicating that synthesis is subject to induction by the substrate. On a growth medium containing sucrose, production was maximal at 15 h in cultures incubated at 28 C°. After this time the level of detectable invertase in the cultures declined. A proportion of the enzyme was secreted during the linear growth phase of the fungus. Various sugars were investigated for induction of invertase, but only the two β-fructofuranosides induced high production levels; with the other sugars, the enzyme was produced only at a low constitutive level. Mycelium grown under repressive conditions (1% glucose), rapidly produced invertase when transferred to sucrose-containing medium. After 80 min the invertase level in these cultures was 26-fold higher than the constitutive level. The repressive effect of other sugars, e.g. glucose and xylose, on invertase production was also demonstrated in this experimental system.
SUMMARY: Protein kinase activities have been detected in cell-free extracts of the cyanobacterium Anabaena PCC 7120. At least 12 polypeptides in the soluble fraction were phosphorylated in vitro at the expense of [y -32P]ATP and the pattern of phosphorylation was shown to be regulated by intermediary metabolites and other effectors, at physiological concentrations. Glucose 6-phosphate exerted a regulatory effect on a phosphopolypeptide of M r 56000 (p56) by stimulating a protein phosphatase, whereas ribulose 5-phosphate inhibited the corresponding protein kinase. In addition, DTT and the calmodulin antagonist trifluoperazine influenced the phosphorylation state of several different polypeptides, indicative of control by redox conditions and a calmodulin-like mediator, respectively. Furthermore, it was established that the phosphorylation of p56 required Mg2+ (> 100 μM) whereas that of a polypeptide of M r 16000 occurred in the absence of Mg2+ and was inhibited by high concentrations (> 1 mM) of this cation. Several of the phosphopolypeptides detected in vitro corresponded in mobility on SDS-PAGE to species phosphorylated in vivo.
SUMMARY: Addition of glucose or related fermentable sugars to derepressed cells of the yeast Saccharomyces cerevisiae triggers a RAS-protein-mediated cAMP signal, which induces a protein phosphorylation cascade. Yeast strains without a functional CDC25 gene were deficient in basal cAMP synthesis and in the glucose-induced cAMP signal. Addition of dinitrophenol, which in wild-type strains strongly stimulates in vivo cAMP synthesis by lowering intracellular pH, did not enhance the cAMP level. cdc25 disruption mutants, in which the basal cAMP level was restored by the RAS2 val19 oncogene or by disruption of the gene (PDE2) coding for the high-affinity phosphodiesterase, were still deficient in the glucose- and acidification-induced cAMP responses. These results indicate that the CDC25 gene product is required not only for basal cAMP synthesis in yeast but also for specific activation of cAMP synthesis by the signal transmission pathway leading from glucose to adenyl cyclase. They also show that intracellular acidification stimulates the pathway at or upstream of the CDC25 protein. When shifted to the restrictive temperature, cells with the temperature sensitive cdc25-5 mutation lost their cAMP content within a few minutes. After prolonged incubation at the restrictive temperature, cells with this mutation, and also those with the temperature sensitive cdc25-1 mutation, arrested at the ‘start’ point (in G1) of the cell cycle, and subsequently accumulated in the resting state G0. In contrast with cdc25-5 cells, however, the cAMP level did not decrease and normal glucose- and acidification-induced cAMP responses were observed when cdc25-1 cells were shifted to the restrictive temperature. These results show that, in the original genetic background at least, growth arrest of cdc25-1-bearing cells at the restrictive temperature is not due to cAMP deficiency. Previous experiments have provided evidence for the presence of a glucose-repressible protein in the signalling pathway. Exponential-phase glucose-grown cells of a strain with overexpression of CDC25 unexpectedly showed a glucose-induced cAMP signal. Control experiments, however, indicated that overexpression of CDC25 caused a defect in glucose repression. Introduction of the cat1 derepression mutation in the strain with overexpression of CDC25 restored glucose repression and abolished the glucose-induced cAMP signal, indicating that the CDC25 protein is not the glucose-repressible component of the signalling pathway.
SUMMARY: Myxococcus xanthus, when starved on a solid surface, undergoes a multicellular cycle of development that consists of fruiting and sporulation. Myxospore germination has been followed in CTT, a complex medium composed mainly of peptides, by monitoring the sequential disappearance of some characteristic spore properties. Loss of heat resistance, shortly after incubation was initiated, was followed by loss of resistance to SDS and finally, refractility and ovoid shape. Germination of a population of myxospores did not occur synchronously. However, in the presence of calcium, germination was activated, being more rapid and synchronous. Other spore activation treatments tested did not have the same stimulatory effect. We searched for specific nutrients or chemicals capable of inducing germination. Amino acids, unlike other potential carbon and energy sources for M. xanthus, or several sugars tested, were most effective in triggering germination. Continuous incubation in CTT and Casamino acids germinant solutions was not required for induction and completion of germination of a large proportion of spores in a population.
SUMMARY: Growth of Synechocystis PCC 6803 in BG-11 medium supplemented with 1 mM-CsCl resulted in intracellular accumulation of Cs+ to a final level of approximately 510 nmol (109 cells)-1 after incubation for 10 d. The doubling time was increased by 64% and the final cell yield was decreased by 70% during growth in the presence of Cs+ as compared to growth in control BG-11 medium. When the total monovalent cation concentration of the medium was doubled by adding either K+ or Na+, levels of accumulated Cs+ were decreased by approximately 50% to 220 and 270 nmol (109 cells)-1, respectively, after 28 d with little inhibition of growth being apparent. Short-term experiments revealed that extracellular K+ and Na+ inhibited Cs+ accumulation to a similar extent, with 90% inhibition of Cs+ accumulation occurring at the highest concentrations used (50 mM-K+ or Na+; 1 mM-Cs+). In all experiments, Cs+ accumulation resulted in a reduction in intracellular K+, except when cells were grown in K+-depleted medium, although a stoichiometric relationship was not apparent, the amount of Cs+ accumulated generally being greater than the amount of K+ released. Cs+ accumulation had no discernible effect on intracellular Na+. When K+, Na+, Rb+, Li+ or Tl+ were supplied at equimolar (1 mM) concentrations to Cs+, only Tl+ significantly reduced Cs+ accumulation. However, an approximately 50% inhibition of Cs+ accumulation resulted when concentrations of K+, Na+, Rb+ or Li+ were increased to 10 mM, which suggests that Cs+ may have a higher affinity for the monovalent cation transport system than K+, Rb+ and TI+ also caused a decrease in intracellular K+, whereas Na+ and Li+ stimulated K+ uptake. Cs+ accumulation was dependent on the external Cs+ concentration and showed a linear relationship to external Cs+ concentrations≤2 mM over 12 h incubation. However, prolonged incubation in external Cs+ concentrations≥ 0·8 mM resulted in Cs+ release from the cells and after 48 h, similar amounts of Cs+ and K+ were present in cells incubated at these higher concentrations. Cs+ accumulation was energy- and pH-dependent. Incubation in the light at 4 °C, or in the presence of 3(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), or at 22 °C in the dark resulted in decreased Cs+ accumulation and decreased K+ release from the cells. Increased amounts of Cs+ were accumulated as the pH of the external medium was increased, with maximal accumulation [approximately 1330 nmol Cs+ (109 cells)-1 after 24 h incubation] occurring at pH 10, the highest pH value used. It is suggested that an important mechanism of Cs+ toxicity in Synechocystis PCC 6803 arises through replacement of cellular K+ by Cs+. The possible role of primary producers such as cyanobacteria in the mobilization of this radionuclide in aquatic habitats is discussed.