Changes in nucleotide pools during conidial germination in Neurospora crassa

1981 ◽  
Vol 59 (11-12) ◽  
pp. 899-905 ◽  
Author(s):  
Richard L. Sabina ◽  
Paulette Dalke ◽  
Alan R. Hanks ◽  
Jane M. Magill ◽  
Clint W. Magill

The acid-soluble nucleotide pools of wild type and several adenine auxotrophs of Neurospora crassa were studied immediately prior to and during conidial germination in the presence of adenine. A two- to four-fold increase in most nucleotide pools was observed after 6 h of germination at 33 °C indicating a general increase in nucleotide pools during this developmental period. The largest components of the acid-soluble nucleotide pools were uracil-containing nucleotide–sugars, which are precursors of chitin and glucan, the major constituents of cell walls. On removal of adenine, the UDP–sugar pools decreased significantly, in adenine auxotrophs, while the pools of UTP increased significantly. ATP levels increased approximately twofold in the first 6 h of germination. After 1 h without exogenous adenine, ATP dropped twofold or more in adenine auxotrophs but not in wild type. There was a net decrease in all adenine nucleotide pools during adenine starvation and a much greater decrease was seen in adenine auxotrophs than in wild type. The adenylate energy charge remained stable despite major changes in the adenylate pools.Accumulation of intermediates was observed in germinated conidia from purine auxotrophs blocked at various steps in the purine pathway. IMP accumulated in ungerminated and in starved conidia of the adenine-8 (ad-8) strain. Ungerminated conidia of the ad-5 strain contained a large pool of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) while the ad-1 strain had smaller amounts of AICAR, but significantly more than any other strain tested. The AICAR pools disappeared from ad-5 and ad-1 in the presence of 50 mg histidine/100 mL. Similarly the IMP pools in ad-8 decreased markedly in the presence of histidine, indicating that the contribution from the histidine biosynthetic pathway to purine nucleotide formation is significant.

Blood ◽  
1977 ◽  
Vol 49 (1) ◽  
pp. 89-99 ◽  
Author(s):  
HJ Reimers ◽  
MA Packham ◽  
JF Mustard

Abstract In rabbit platelets, the metabolically active ATP pool equilibrates with the releasable ATP pool within 1 day. The studies showing this have now been extended to human platelets. Human platelets labeled with 14C-adenosine or 14C-adenine were incubated for up to 10 hr in vitro at 37 degrees C. After 10 hr, about 12% of the total platelet 14C-ATP and 14C-ADP had become releasable with thrombin (4.2 units/ml). Lysis of platelets did not occur, since less than 1% of the platelet-bound 51Cr from platelets labeled with this radioisotope appeared in the ambient fluid upon thrombin treatment. The 14C-ATP/14C-ADP ratio of the released adenine nucleotides (7.6) was similar to the 14C-ATP/14C-ADP ratio of the nonreleasable adenine nucleotides (7.1) 2 hr after the labeling with 14C-adenosine. However, upon prolonged incubation (10 hr) in vitro, the 14C-ATP/14C-ADP ratio of the releasable adenine nucleotides decreased to 2.7. The adenylate energy charge and the 14C- ATP/14C-ADP ratio of the metabolic adenine nucleotide pool did not change significantly during the time of observation. The 14C-ATP content of the platelets decreased by less than 1% hr of incubation at 37 degrees C. These observations are interpreted to mean that the 14C is transferred from the metabolically active, nonreleasable adenine nucleotide pool of human platelets into the releasable adenine nucleotide pool as ATP and is partially hydrolyzed there to yield ADP. The transfer of ATP across the storage organelle membrane of platelets may be similar to transport processes in the chromaffin cells of the adrenal medulla and may represent a general phenomenon in cells that possess storage organelles containing adenine nucleotides.


1984 ◽  
Vol 224 (3) ◽  
pp. 961-969 ◽  
Author(s):  
T D Paul ◽  
P W Ludden

Adenine nucleotide pools were measured in Rhodospirillum rubrum cultures that contained nitrogenase. The average energy charge [([ATP] + 1/2[ADP])/([ATP] + [ADP] + [AMP])] was found to be 0.66 and 0.62 in glutamate-grown and N-limited cultures respectively. Treatment of glutamate-grown cells with darkness, ammonia, glutamine, carbonyl cyanide m-chlorophenylhydrazone, or phenazine methosulphate resulted in perturbations in the adenine nucleotide pools, and led to loss of whole-cell nitrogenase activity and modification in vivo of the Fe protein. Treatment of N-limited cells resulted in similar changes in adenine nucleotide pools but not enzyme modification. No correlations were found between changes in adenine nucleotide pools or ratios of these pools and switch-off of nitrogenase activity by Fe protein modification in vivo. Phenazine methosulphate inhibited whole-cell activity at low concentrations. The effect on nitrogenase activity was apparently independent of Fe protein modification.


1976 ◽  
Vol 54 (6) ◽  
pp. 886-891 ◽  
Author(s):  
J. H. A. Fields ◽  
H. Guderley ◽  
K. B. Storey ◽  
P. W. Hochachka

Citrate synthase (EC 4.1.3.7) in mantle muscle of the octopus, Octopus cyanea, occurs in relatively low specific activity and is largely independent of pH between 7.5 and 9.0. Catalytic activity is regulated by the adenylate energy charge and by at least two Krebs cycle intermediates, α-ketoglutarate and citrate. Of the adenylates, ATP is by far the most potent inhibitor, at near-physiological concentrations (4 mM), causing almost a 20-fold increase in the Michaelis constant for acetyl-CoA. Citrate and α-ketoglutarate, on the other hand, are competitive with respect to oxaloacetate, rather than acetyl-CoA, and bring about large increases in the Michaelis constant for oxaloacetate. The regulatory properties of citrate synthase allow a curtailment of carbon flow into the Krebs cycle during periods of burst muscle work, when mantle anaerobic glycolysis is strongly activated.


Parasitology ◽  
1980 ◽  
Vol 81 (3) ◽  
pp. 593-601 ◽  
Author(s):  
M. J. Sharpe

SUMMARYThe adenine nucleotide content and adenylate energy charge ofNematospiroides dubiusfrom laboratory mice and ofTrichostrongylus colubriformisfrom lambs has been measured. Administration of the anthelmintic, levamisole, to infected hosts resulted in only a slight fall in the adenylate energy charge ofN. dubiusover a 3-h period but there was a greater fall in the adenylate energy charge ofT. colubriformisduring this period. In neither case did the energy charge fall quickly, nor did it fall to the low levels which would be expected if the levamisole were inhibiting synthesis of ATP. The changes in energy charge of the nematodes which occurred following administration of levamisole to their hosts was of the order which can be satisfactorily explained by changes in the environment of the nematodes, such as reduced oxygen tension. It is concluded that the maintenance of levamisole-induced paralysis of these two species of trichostrongylein vivodoes not rely on the inhibition of fumarate reductase.


Blood ◽  
1981 ◽  
Vol 57 (2) ◽  
pp. 203-208
Author(s):  
AK Rao ◽  
S Niewiarowski ◽  
S Murphy

Platelets stored as concentrates (PC) for 72 h at 22 degrees C develop a functional defect. Alterations in adenine nucleotides of platelets have been shown to affect platelet function. Adenine nucleotide content of platelets was measured before and after storage and a decrease of 27.1 /+- 1.7% (mean /+- SE) in ATP and 39.1 /+- 2.6% in ADP were found in 34 PC stored with final volume of 50 ml. In 11 PC with 30 ml volume. ATP and ADP decreased by 39.4 /+- 3.2% and 49.4 /+- 2.1%, respectively. The mean ATP to ADP ratio of stored platelets was significantly higher than of fresh platelets in both groups, suggesting a relatively greater decrease in granular than metabolic pool nucleotides. Levels of low affinity platelet factor 4 measured by radioimmunoassay in plasma from 0.86 /+- 0.08 microgram/ml in the fresh PC to 8.59 /+- 0.39 microgram/ml in stored PC, indicating a concomitant alpha-granular secretion. Labeling of metabolic pool with 14C-adenine revealed a mean decrease in the adenylate energy charge of 2.0 /+- 0.4% in 12 of 16 stored PC, with a lower ATP and higher hypoxanthine labeling in stored as compared to fresh platelets. These observations suggest that stored platelets develop an acquired defect in both dense and alpha granules and in their ability to maintain ATP homeostasis.


HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 842A-842
Author(s):  
Shahid N. Chohan ◽  
Terence A. Brown

HPLC was used to measure nucleotide pools in tomato seeds during the 3 h following imbibition in water. In dry seeds, nucleotides were predominantly in the form of monophosphates. During the first 2 h post-imbibition, the monophosphate levels declined and there were sharp increases in the amounts of diphosphates, followed by triphosphates. Between the 2nd and 3rd h, the di- and triphosphate levels continued to increase and the monophosphate levels began to recover, especially in the case of UMP—presumably the result of degradation of unwanted mRNAs left over from the maturation phase of seed development. The adenylate energy charge increased during imbibition and, within 3 h, reached a value close to that of normal active tissue.


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