Tryptose phosphate broth confers to chick embryo cells resistance to the inhibitory effect of chloramphenicol on growth

Leblond-Larouche, Lorraine; Larouche, Alexandrine; Guertin, Denise; Morais, Réjean

doi:10.1016/0006-291x(77)91614-x

Cited by 14 publications

(6 citation statements)

References 21 publications

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“…Evidence for this has been mainly documented by studies showing that specific inhibitors of mitochondrial DNA replication, transcription, and translation limited animal cell growth to a few generations (1)(2)(3)(4)(5). The cessation of growth has been generally believed to result from an insufficient energy production following the gradual inhibition of the mitochondrial respiratory capacity of the cell.…”

Section: Introductionmentioning

confidence: 96%

“…Insight into the contribution of the mitochondrial genome to the growth of animal cells was gained when we observed that respiration-deficient chick embryo cells are inherently resistant to the growth inhibitory effect of chloramphenicol when cultured in the presence of tryptose phosphate broth (4,13) or its active components of pyrimidine origin (15,16). Further studies indicated that auxotrophy for pyrimidines appears to result from a deficiency in dihydroorotate dehydrogenase activity (M. Grdgoire, R. Morais, D. Gravel, and M. A. Quilliam, in preparation), a mitochondrially located enzyme (17).…”

Section: Introductionmentioning

confidence: 98%

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On the contribution of the mitochondrial genome to the growth of Chinese hamster embryo cells in culture

Morais¹,

Guertin²,

Kornblatt³

1982

Can. J. Biochem.

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The present results demonstrate that Chinese hamster embryo cell populations in culture can be adapted to grow in the presence of chloramphenicol. It is shown that tryptose phosphate broth and uridine, one of its components, prevent the growth inhibitory effect of the drug. Study of some respiratory parameters (cytochrome c oxidase, cytochrome spectra, oxygen consumption) indicated that neither the broth nor uridine prevented the inhibitory effect of chloramphenicol on mitoribosomal protein synthesis. The cells grew with mitochondria devoid of a functional respiratory chain. Auxotrophy for pyrimidines appeared to result from the absence of dihydroorotate dehydrogenase, a respiratory chain-linked enzyme that catalyzes the fourth step of de novo pyrimidine biosynthesis. These and other results suggest that the synthesis of orotic acid may be considered as one of the main contributions of mitochondria to the growth of animal cells in culture.

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Section: Introductionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 98%

On the contribution of the mitochondrial genome to the growth of Chinese hamster embryo cells in culture

Morais¹,

Guertin²,

Kornblatt³

1982

Can. J. Biochem.

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“…This enzyme is also required for the electron transport chain (Chen & Jones, 1976). In addition, TPB abrogates the inhibitory effect of chloramphenicol on the growth of chick embryo fibroblasts (Leblond-Larouche et al, 1977). Because of this inhibitory effect, the decrease in mitochondrial ATP production appears to be compensated for by an increase in the activities of pyruvate kinase and lactate dehydrogenase (Leblond-Larouche et al, 1977).…”

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confidence: 99%

Tryptose phosphate broth improvesRickettsia felisreplication in mammalian cells: 1

Saisongkorh

Karkouri

Patrice

et al. 2012

FEMS Immunol Med Microbiol

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In cell culture, Rickettsia felis grows only at low temperatures (< 31 °C). Therefore, its ability to enter, survive and grow in cell lines has primarily been tested in cells derived from amphibians and arthropods, which naturally grow at low temperatures, and only infrequently in mammalian cells. We subcultured R. felis in mammalian cells for more than 10 passages using media supplemented with tryptose phosphate broth (TPB) and found that TPB is critical for optimal growth of R. felis in mammalian cells.

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“…We previously reported [1,2] that the growth of chick embryo cell populations (CEF), treated with chloramphenicol, ceases almost completely after a few generations. Table 1 summarizes the results obtained from numerous experiments.…”

Section: Initial Observationsmentioning

confidence: 99%

On auxotrophy for pyrimidines of respiration‐deficient chick embryo cells

Gregoire

Morais

Quilliam

et al. 1984

European Journal of Biochemistry

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Chick embryo cells treated with chloramphenicol are inherently resistant to the growth-inhibitory effect of the drug when cultured in the presence of tryptose phosphate broth. The cells were found to be auxotrophic for pyrimidines and the presence in the broth of compounds of pyrimidine origin is demonstrated by chromatographic procedures and mass spectral analyses. They are in the form of ribonucleosides, ribonucleotides and pyrimidinecontaining oligoribonucleotides. To understand the mechanism responsible for pyrimidine auxotrophy, the activity of enzymes involved in the pyrimidine biosynthetic pathway was determined. Measurement of the conversion of dihydroorotic acid to orotic acid in cell-free extracts revealed that chloramphenicol-treated chick embryo cells are deficient in dihydroorotate dehydrogenase activity. The data in vitro are supported by studies on the nutritional requirements of the respiration-deficient cells and by the incorporation in vivo of labelled dihydroorotic acid into the acid-insoluble fraction of the cells. Although the activity of the dehydrogenase in vitro is decreased by 95 %, the enzyme is present in chlor amphenicol-treated cells and its activity is unmasked by the artificial electron acceptor menadione. A study of the activity of other enzymes of the pyrimidine biosynthetic pathway demonstrated that their activity is comparable to that in control cells. The present results indicate that auxotrophy for pyrimidines results from the inhibition of the flow of electrons along the mitochondrial electron transport chain.Previous work from this laboratory has demonstrated that chick embryo cell populations are inherently resistant to the growth-inhibitory effect of chloramphenicol and ethidium bromide (EtdBr) when the culture medium is supplemented with tryptose phosphate broth [l -31. The broth is an enzymatic digest of animal extracts supplemented with dextrose, sodium chloride and disodium phosphate [4]. Study of growth parameters indicated that no lag or adaptation period appeared necessary for the broth-treated chick cell populations to proliferate in the presence of the drugs. Furthermore, measurements of mitochondrial respiratory parameters demonstrated that the broth does not prevent the inhibitory effect of chloramphenicol and EtdBr on the mitochondrial marcromolecular synthesizing systems [I -31. The cells grow with mitochondria devoid of a functional respiratory chain.In the present paper it is demonstrated that the active components of the broth are of pyrimidine origin in the form of ribonucleosides, ribonucleotides and pyrimidine-containing oligoribonucleotides. In addition, a study of the activity of different enzymes involved in de novo biosynthesis of uridylic acid is included. Auxotrophy for pyrimidines appears to result from a deficiency in dihydroorotate dehydrogenase activity, an enzyme located in the mitochondria.

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Tryptose phosphate broth confers to chick embryo cells resistance to the inhibitory effect of chloramphenicol on growth

Cited by 14 publications

References 21 publications

On the contribution of the mitochondrial genome to the growth of Chinese hamster embryo cells in culture

On the contribution of the mitochondrial genome to the growth of Chinese hamster embryo cells in culture

Tryptose phosphate broth improvesRickettsia felisreplication in mammalian cells: 1

On auxotrophy for pyrimidines of respiration‐deficient chick embryo cells

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