The metabolism of pyrimidine compounds by Tetrahymena pyriformis

The pyrimidine requirements for growth of T. pyriformis and for reversal of the growth inhibition caused by folate deprivation have been studied. The effects of thymidine and 5-fluorodeoxyuridine have been shown to be quantitatively different from the effects of these compounds on growth and the rate of DNA synthesis in mammalian cells. Labelled nucleosides added to the medium have been found to be converted to the corresponding bases with the exception of deoxycytidine, which is first deaminated to deoxyuridine. As a result no deoxynucleosides other than thymidine specifically label DNA. The results allow deductions to be made concerning the enzymes involved in pyrimidine utilization by this organism. It is suggested that pyrimidine utilization is always channeled through uracil in the case of those compounds that can supply the pyrimidine requirement for growth.     

The purine and pyrimidine metabolism of Tetrahymena pyriformis

The metabolism of purines and pyrimidines by the ciliated protozoan Tetrahymena was investigated with the use of enzymatic assays and radioactive tracers. A survey of enzymes involved in purine metabolism revealed that the activities of inosine and guanosine phosphorylase (purine nucleoside: orthophosphate ribosyltransferase, E.C. 2.4.2.1) were high, but adenosine phosphorylase activity could not be demonstrated. The apparent K_m for guanosine in the system catalyzing its phosphorolysis was 4.1 ± 0.6 × 10^?3 M. Pyrophosphorylase activities for IMP and GMP (GMP: pyrophosphate phosphoribosyltransferase, E.C. 2.4.2.8), AMP (AMP: pyrophosphate phosphoribosyltransferase, E.C. 2.4.2.7), and 6-mercaptopurine ribonucleotide were also found in this organism; but a number of purine and pyrimidine analogs did not function as substrates for these enzymes. The metabolism of labeled guanine and hypoxanthine by intact cells was consistent with the presence of the phosphorylases and pyrophosphorylases of purine metabolism found by enzymatic studies. Assays for adenosine kinase (ATP: adenosine 5'-phosphotransferase, E.C. 2.7.1.20) inosine kinase, guanosine kinase, xanthine oxidase (xanthine: O₂ oxidoreductase, E.C. 1.2.3.2), and GMP reductase (reduced-NADP: GMP oxidoreductase [deaminating], E.C. 1.6.6.8) were all negative. In pyrimidine metabolism, cytidine-deoxycytidine deaminase (cytidine aminohydrolase, E.C. 3.5.4.5), thymidine phosphorylase (thymidine: orthophosphate ribosyltransferase, E.C. 2.4.2.4), and uridine-deoxyuridine phosphorylase (uridine: orthophosphate ribosyltransferase, E.C. 2.4.2.3) were active; but cytidine kinase, uridine kinase (ATP: uridine 5'-phosphotransferase, E.C. 2.7.1.48), and CMP pyrophosphorylase could not be demonstrated.     

The effects of supraoptimal temperatures on population growth and cortical patterning in Tetrahymena pyriformis and Tetrahymena thermophila: a comparison

In this investigation, we compare the multiplication rates and morphogenetic responses of the two most studied Tetrahymena species, T. pyriformis and T. thermophila, at supraoptimal temperatures. Although the upper temperature limits differ greatly in the two species, the pattern of growth responses to high temperature is for the most part similar, with some differences in detail. The transient recovery of cell division at the highest temperature that allows cell division, characteristic of T. pyriformis, is observed in a less distinct form in T. thermophila. Moreover, there is a remarkable difference in developmental response, with drastic abnormalities in patterning of oral structures during the transient recovery of cell division in T. pyriformis, and far more limited abnormalities under similar conditions in T. thermophila. The abnormalities result from spatial disorder in the alignment and orientation of basal body pairs within the early oral primordium, followed by failures in the realignment that normally occurs as oral structures (membranelles and undulating membrane) mature. Both the initial spatial disorder and the failures in realignment are far more severe in T. pyriformis than in T. thermophila.     

Steric effects at C-20 and C-24 on the metabolism of sterols by Tetrahymena pyriformis

Cultures of Tetrahymena pyriformis were incubated with various sterols and the extent of dehydrogenation at C-7 and C-22 was determined. The sterols incubated were desmosterol, 22-dehydrodesmosterol, 24-methyldesmosterol, 24 alpha-methylcholesterol (campesterol), 24-methylene-cholesterol, isohalosterol (26,27-bisnorcampesterol, also known as 24,24-dimethylchol-5-en-e beta-ol, a naturally occurring C26-sterol), and 20-isohalosterol. 20-Isohalosterol was not metabolized, while products with delta 7- and delta 22-bonds were formed from isohalosterol and all of the other sterols studied. This confirms an earlier conclusion, based on results with 20-isocholesterol and cholesterol, that inversion of the configuration from 20(R) to 20(S) completely prevents metabolism both in the nucleus and the side chain. On the other hand, changes in the electronics or stereochemistry at C-24 had a direct affect only on metabolism in the side chain. The presence of a methyl group at C-24 reduced the yield of metabolites with a delta 22-bond relative to those with a delta 7-bond producing an accumulation of 7-dehydro metabolite. A double bond at position-24 counteracted this steric effect, presumably by enhancing the rate of dehydrogenation, and a delta 24(28)-bond was more effect than was a delta 24(25)-bond.     

Influence of ions and tonicity of RNA metabolism in Tetrahymena pyriformis

Net RNA degradation occurs in Tetrahymem pyrifmmis when this ciliate is suspended in a non-nutrient medium. The quantity and quality of the excretion products is at least partially under the control of the ionic content and the tonicity of the cellular environment. The excretion of ultraviolet-absorbing materials was found to be elevated by sodium ions in a medium isotonic to the culture fluid, or by a hypertonic environment. Magnesium counteracted these effects. In isotonic suspension, sodium and magnesium ions lowered orthophosphate excretion; however, sodium altered the nature of the phosphate products so that acidlabile phosphates were also excreted rather than solely orthophosphate. Similar results were obtained in a hypertonic environment with or without sodium. The degree of purine and pyrimidine loss from the cells in all conditions of suspension was reflected in the amount of RNA degraded. The ion and tonicity effects apparently reflect events which alter the stability of the RNA and the properties of the membrane system, resulting in changes in both the rate of RNA degradation and the nature of the excreted products. The rates of orthophosphate excretion appear to be affected by changes in the acid-base balance within the cell which may be governed by the cation levels. The manipulation of the ionic content and tonicity of the medium offers a convenient method for obtaining cells reduced in RNA content.