scholarly journals NUCLEOLAR AND BIOCHEMICAL CHANGES DURING UNBALANCED GROWTH OF TETRAHYMENA PYRIFORMIS

1965 ◽  
Vol 26 (3) ◽  
pp. 845-855 ◽  
Author(s):  
Ivan L. Cameron ◽  
E. Ernest Guile
Keyword(s):  
Protein Synthesis ◽  
Stationary Phase ◽  
Rna Synthesis ◽  
Morphological Changes ◽  
Synthetic Medium ◽  
Tetrahymena Pyriformis ◽  
Lag Phase ◽  
Proteose Peptone ◽  
Rna And Protein Synthesis ◽  
Peptone Medium

Numerous nucleoli can be observed in the macronucleus of the logarithmically growing ciliated protozoan Tetrahymena pyriformis; at late log phase the nucleoli aggregate and fuse. In stationary phase this fusion process continues, leaving a very few large vacuolated nuclear fusion bodies in the nucleus. When these stationary phase cells are placed into fresh enriched proteose peptone medium, the large fusion bodies begin to disaggregate during the 2.5-hour lag phase before cell division is initiated. By 3 to 6 hours after inoculation the appearance of the nucleoli in many cells returns to what it was in logarithmic cells. In view of the possible role of nucleoli in ribosome synthesis, attempts were made to correlate the morphological changes to changes in RNA and protein metabolism. The beginning of an increased RNA synthesis was concomitant with the beginning of disaggregation of the large fusion bodies into nucleoli, which was noticed in some cells by 1 hour after the return to fresh enriched proteose peptone medium. Increased protein synthesis then followed the increased RNA synthesis by 1 hour. The supply of RNA precursors (essential pyrimidines) were removed from cultures which were grown on a chemically defined synthetic medium, in order to study the relation between nucleolar fusion and synthesis of RNA and protein. Pyrimidine deprivation drastically curtailed RNA and protein synthesis, but did not cause fusion of nucleoli. When pyrimidines were added back to this culture medium, RNA synthesis was immediately stimulated and again preceded an increased protein synthesis by 1 hour. These studies suggest the involvement of unfused nucleoli in RNA and protein synthesis and demonstrate the extreme plasticity of nucleoli with respect to changes in their environment.

Regulation of RNA synthesis in Tetrahymena pyriformis: secretion of regulatory factors

1979 ◽  
Vol 35 (1) ◽  
pp. 17-24
Author(s):  
H.A. Andersen ◽  
S.J. Nielsen
Keyword(s):  
Stationary Phase ◽  
Ribosomal Rna ◽  
Rna Synthesis ◽  
Initial Rate ◽  
Tetrahymena Pyriformis ◽  
Dose Effect ◽  
Exponential Growth Phase ◽  
Lag Phase ◽  
Cell Densities ◽  
Phase Population

The rate of ribosomal RNA synthesis varies greatly with the population density in both exponentially and synchronously growing populations of Tetrahymena pyriformis. Shortly after inoculation of the population - at relatively low cell densities - a gene-dose effect dominates the picture, and a doubling in the gene number is immediately followed by a doubling in the rate of RNA synthesis. However, also other mechanisms are controlling the rate of RNA synthesis. Generally one finds high rates of RNA synthesis in the lag phase of newly inoculated cells, decreasing rate of RNA synthesis during most of the exponential growth phase and very low rate of synthesis in stationary phase cells. We now have results which show that the repression of RNA synthesis in densely populated cultures is caused by a dialysable factor, which is secreted by the cells. If cells are inoculated on a medium which contains this factor the high initial rate of RNA synthesis normally observed is prevented, but the cells multiply and grow with normal generation time until normal stationary-phase population densities are reached.

scholarly journals Changes in the transport of mitochondrial Ca2+ during the culture growth cycle of Tetrahymena pyriformis

1985 ◽  
Vol 77 (1) ◽  
pp. 47-56
Author(s):  
Y.V. Kim ◽  
LYu Kudzina ◽  
V.P. Zinchenko ◽  
Y.V. Evtodienko
Keyword(s):  
Stationary Phase ◽  
Morphological Changes ◽  
Stationary Phases ◽  
Mitochondrial Protein ◽  
Tetrahymena Pyriformis ◽  
Growth Cycle ◽  
Lag Phase ◽  
Rapid Release ◽  
Culture Growth ◽  
Energy Dependent

The properties of the Ca2+ transport system of mitochondria, isolated in various phases of growth of static cultures of Tetrahymena pyriformis, were studied. A large increase in the endogenous energy-dependent Ca2+ content of mitochondria was observed as cultures of T. pyriformis passed through the exponential and stationary phases of growth (approx. 0.25 and 50 nmol Ca2+ per mg mitochondrial protein, respectively). Simultaneously, the mitochondria dramatically lost their ability to withstand large concentrations of Ca2+ and ADP. However, in the latter case they were able to phosphorylate a large amount of ADP if the strong Ca2+ chelator, ethylene glycol bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, was initially present in the incubation medium. Furthermore, all the changes observed in mitochondria from the stationary phase cells were completely reversed when cell proliferation was re-activated after the lag phase, either by reseeding the stationery cells in fresh growth medium or by oxygenation of the old medium. In aerobic conditions even a small addition of Ca2+ was able to induce rapid release of Ca2+ from mitochondria isolated during the stationary phase of growth. It is suggested that the redistribution of Ca2+ between the mitochondria and the cytoplasm at the onset of the lag phase may serve as the main trigger for the subsequent biochemical and morphological changes observed in T. pyriformis.

FEMALE ENDOCRINE CONTROL MECHANISMS DURING THE NEONATAL PERIOD

1972 ◽  
Vol 70 (2) ◽  
pp. 396-408 ◽  
Author(s):  
K.-D. Schulz ◽  
H. Haarmann ◽  
A. Harland
Keyword(s):  
Protein Synthesis ◽  
Reproductive System ◽  
Rna Synthesis ◽  
Neonatal Period ◽  
High Dose ◽  
Female Reproductive System ◽  
Endocrine Control ◽  
Hypothalamic Region ◽  
Rna And Protein Synthesis ◽  
Physiological Dose

ABSTRACT The present investigation deals with the oestrogen-sensitivity of the female reproductive system during the neonatal period. Newborn female guinea pigs were used as test animals. At different times after a single subcutaneous injection of a physiological dose of 0.1 μg or an unphysiologically high dose of 10 μg 17β-oestradiol/100 g body weight, the RNA- and protein-synthesis was examined in the hypothalamic region, pituitary, cerebral cortex, liver, adrenal gland, ovary and uterus. With a physiological dose an increase in organ weight, protein content, RNA-and protein-synthesis was found only in the uterus. These alterations turned out to be dose-dependent. In addition to the findings in the uterus an inhibition of the aminoacid incorporation rate occurred in the liver following the injection of the high oestradiol dose. As early as 1 hour after the administration of 0.1 μg 17β-oestradiol an almost 100% increase in uterine protein synthesis was detectable. This result demonstrates a high oestrogen-sensitivity of this organ during the neonatal period. All the other organs of the female reproductive system such as the hypothalamus, pituitary and ovary did not show any oestrogen response. Therefore the functional immaturity of the uterus during post partem life is not the result of a deficient hormone sensitivity but is correlated with the absence of a sufficient hormonal stimulus at this time. The investigation on the effects of actinomycin resulted in different reactions in the uterus and liver. In contrast to the liver a paradoxical actinomycin effect was found in the uterus after treatment with actinomycin alone. This effect is characterized by a small inhibition of RNA-synthesis and a 50% increase in protein synthesis. The treatment of the newborn test animals with actinomycin and 17β-oestradiol together abolished the oestrogen-induced stimulation of the uterine RNA-and protein-synthesis. Consequently, the effect of oestrogens during the neonatal period is also connected with the formation of new proteins via an increased DNA-directed RNA-synthesis.

Die Bildung von Thymidintriphosphat bei Tetrahymena pyriformis (EHRENBERG) in statistischen und hitzesynchronisierten Kulturen / Formation of Thymidine Triphosphate in statistical and heat synchronized cultures of Tetrahymena pyriformis (EHRENBERG)

1970 ◽  
Vol 25 (5) ◽  
pp. 517-521 ◽  
Author(s):  
Manfred K. Grieshaber ◽  
Franz Duspiva
Keyword(s):  
Heat Treatment ◽  
Enzyme Activity ◽  
Stationary Phase ◽  
Dna Synthesis ◽  
Tetrahymena Pyriformis ◽  
Thymidylate Kinase ◽  
Proteose Peptone ◽  
Thymidine Triphosphate ◽  
Logarithmic Growth ◽  
Synchronized Cultures

The activity of thymidylate kinase is correlated with DNA-synthesis in Tetrahymena. During logarithmic growth it is twice as high as in the stationary phase; in cultures synchronized by heat treatment, the activity of the enzyme increases with the onset of DNA-synthesis. After application of 5 x 10-6 ᴍ Methotrexate, the activity of thymidylate kinase remains unchanged. There is, however, a dramatic increase in enzyme activity when the inhibition of transmethylations is circumvened by adding thymidine and proteose-peptone.

scholarly journals A Synthetic Congener Modeled on a Microbicidal Domain of Thrombin- Induced Platelet Microbicidal Protein 1 Recapitulates Staphylocidal Mechanisms of the Native Molecule

2006 ◽  
Vol 50 (11) ◽  
pp. 3786-3792 ◽  
Author(s):  
Yan Q. Xiong ◽  
Arnold S. Bayer ◽  
Lisa Elazegui ◽  
Michael R. Yeaman
Keyword(s):  
Protein Synthesis ◽  
Rna Synthesis ◽  
Membrane Permeabilization ◽  
Dna And Rna ◽  
Activated Platelets ◽  
Rna And Protein Synthesis ◽  
Dna And Rna Synthesis ◽  
Platelet Microbicidal Protein ◽  
Α Helix ◽  
Native Host

ABSTRACT Thrombin-induced platelet microbicidal protein 1 (tPMP-1) is a staphylocidal peptide released by activated platelets. This peptide initiates its microbicidal activity by membrane permeabilization, with ensuing inhibition of intracellular macromolecular synthesis. RP-1 is a synthetic congener modeled on the C-terminal microbicidal α-helix of tPMP-1. This study compared the staphylocidal mechanisms of RP-1 with those of tPMP-1, focusing on isogenic tPMP-1-susceptible (ISP479C) and -resistant (ISP479R) Staphylococcus aureus strains for the following quantitative evaluations: staphylocidal efficacy; comparative MIC; membrane permeabilization (MP) and depolarization; and DNA, RNA, and protein synthesis. Although the proteins had similar MICs, RP-1 caused significant killing of ISP479C (<50% survival), correlating with extensive MP (>95%) and inhibition of DNA and RNA synthesis (>90%), versus substantially reduced killing of ISP479R (>80% survival), with less MP (55%) and less inhibition of DNA or RNA synthesis (70 to 80%). Interestingly, RP-1-induced protein synthesis inhibition was equivalent in both strains. RP-1 did not depolarize the cell membrane and caused a relatively short postexposure growth inhibition. These data closely parallel those previously reported for tPMP-1 against this strain set and exemplify how synthetic molecules can be engineered to reflect structure-activity relationships of functional domains in native host defense effector molecules.

The relationship of cardiolipin biosynthesis to mitochondrial protein synthesis in Tetrahymena pyriformis

1983 ◽  
Vol 61 (4) ◽  
pp. 223-228 ◽  
Author(s):  
Sean M. Hemmingsen ◽  
Laura Querengesser ◽  
Paul G. Young
Keyword(s):  
Protein Synthesis ◽  
Stationary Phase ◽  
Mitochondrial Protein ◽  
Tetrahymena Pyriformis ◽  
Pulse Labelling ◽  
Mitochondrial Protein Synthesis ◽  
Mole Percent ◽  
Quantitative Measurements ◽  
Relationship Of ◽  
The Relationship

The synthesis of cardiolipin has been investigated following the inhibition of mitochondrial protein synthesis with chloramphenicol. Quantitative measurements of the amount of cardiolipin in the cell during treatment with chloramphenicol as well as pulse-labelling studies using labelled acetate were carried out. The results show that while whole cell phospholipid biosynthesis is depressed by the treatment (probably a reflection of a general cessation of growth), there is no sign of any preferential effect on cardiolipin synthesis. The data also show that as cells are reduced in size as they approach stationary phase there is a six- to seven-fold loss of total cellular phospholipids; however, the amount of cardiolipin is only reduced by four- to five-fold. There is a preferential conservation of cardiolipin as stationary phase is approached with the mole percent cardiolipin phosphorus in the cell rising from 5–7% to 10–12%.

scholarly journals USE OF GENOMIC EXCLUSION TO ISOLATE HEAT-SENSITIVE MUTANTS IN TETRAHYMENA

Genetics ◽  
1973 ◽  
Vol 73 (4) ◽  
pp. 543-559
Author(s):  
Eduardo Orias ◽  
Miriam Flacks
Keyword(s):  
Room Temperature ◽  
Axenic Culture ◽  
Tetrahymena Pyriformis ◽  
Recessive Mutation ◽  
Allelic Exclusion ◽  
Current Interest ◽  
Proteose Peptone ◽  
Abnormal Form ◽  
Mutant Strains ◽  
Peptone Medium

ABSTRACT We have used the abnormal form of conjugation known as "genomic exclusion" to isolate a collection of heat-sensitive mutants of Tetrahymena pyriformis, syngen 1. Growth at room temperature in bacterized medium and no growth at 40°C in the same medium was the criterion used for the isolation. The mutant strains were tested for growth in pure (axenic) culture in proteose peptone medium; of the 31 strains which grew normally at room temperature and not at 40°C in that medium, 21 also failed to grow at 37°C. Preliminary results of complementation tests suggest that most, if not all, the mutations are recessive and that a variety of genes was affected. A detailed genetic analysis was performed on one mutant (H9). The results are all consistent with the idea that the heat-sensitive phenotype of this mutant is determined by a single recessive mutation, designated ts-2. Heterozygotes ts-2/+ yield heat-sensitive segregants during vegetative growth; we interpret this finding as another example of allelic exclusion, a phenomenon universally encountered among heterozygotes in syngen 1 of T. pyriformis. Our results are discussed in the context of some questions of current interest in Tetrahymena genetics.

Metabolic requirements for interactions between nuclear and cytoplasmic membranes in the repair of damaged Amoeba nuclei

1976 ◽  
Vol 21 (2) ◽  
pp. 291-302
Author(s):  
C.J. Flickinger
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Synthesis ◽  
Membrane Fusion ◽  
Energy Production ◽  
Rna Synthesis ◽  
Metabolic Inhibitors ◽  
Protein Synthesis Inhibitors ◽  
Nuclear Membranes ◽  
Cytoplasmic Membranes ◽  
Rna And Protein Synthesis

Amoeba nuclear envelopes were damaged using microsurgery, and metabolic requirements for the steps in their repair were studied, and my placing the cells in a solution containing one of several metabolic inhibitors. The first step in repair, the association of pieces of endoplasmic reticulum with holes in the nuclear membranes, appears to be a passive process since it was not affected by inhibitors of energy production, RNA synthesis, or protein synthesis. In contrast, fusion of pieces of endoplasmic reticulum with the nuclear membranes at the margins of the holes was blocked by KCN and dinitrophenol, indicating that membrane fusion requires energy derived from respiration, but RNA and protein synthesis inhibitors did not prevent fusion of pieces of endoplasmic reticulum with the nuclear membranes. The subsequent completion of repair and restoration of intact nuclear membranes was almost completely blocked by inhibitors of respiration, and it was reduced in the presence of actinomycin and emetine, suggesting that in addition to a requirement for energy, some later steps in the repair of the nuclear membranes require RNA and protein synthesis.

scholarly journals MACROMOLECULAR SYNTHESES RELATED TO THE REPRODUCTIVE CYST OF TETRAHYMENA PATULA

1973 ◽  
Vol 59 (3) ◽  
pp. 615-623 ◽  
Author(s):  
P. R. Gabe ◽  
L. E. de Bault
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Generation Time ◽  
Rna Synthesis ◽  
Culture Medium ◽  
Tritiated Thymidine ◽  
Growth Cycle ◽  
The Third ◽  
Rna And Protein Synthesis ◽  
The Mean

Macromolecular syntheses in encysted Tetrahymena patula were studied using Feulgen fluorescence cytophotometry, autoradiography, and inhibitors of RNA and protein synthesis. Cycloheximide significantly depressed protein synthesis and D-actinomycin effectively blocked RNA synthesis. Under these conditions, the cells within the cyst were unable to divide. Both cytophotometric measurements and autoradiographic data with tritiated thymidine show that DNA synthesis does not occur during the encystment divisions. Excysted cells placed in nutrient broth medium showed a prolonged generation time after the first cell growth cycle, and by the third generation the mean DNA content per cell was almost triple that of starved excysted cells. These findings indicate that (a) the encystment divisions require RNA and protein synthesis, which are apparently effected through turnover, (b) the encystment division cycles occur in the absence of DNA synthesis, and (c) excysted cells placed in culture medium may go through more than one DNA replication per cell cycle.

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