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  • 1
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 268 (1977), S. 64-67 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The mutant cells, V79-G7, were characterised as respiration-deficient by the following criteria8'10. (1) The rate of oxygen consumption by whole cells is about 10% that of wild-type cells. (2) There is no appreciable release of 14COa from 2-14C-pyruvate or 5-14-C-glutamate (conversions requiring an ...
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Somatic cell and molecular genetics 5 (1979), S. 441-451 
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have previously classified 35 of our respiration-deficient mutants into seven complementation groups and one “overlapping” mutant which does not complement mutants from groups I and II. In this paper we report on the biochemical characterization of representatives of complementation groups I, II, VII, and the “overlapping” mutant. We show that these mutants all have a defect in complex I of the electron-transport chain. The general features of these mutants are: (1) a low rate of O2 consumption in whole cells; (2) a low rate of release of14CO2 from [2-14C]pyruvate, [1-14C]pyruvate, and [3-14 C]β-hydroxybutyrate; (3) a low rate of release of14CO2 from [5-14C]glutamate and [1-14C]glutamate in mutants from groups II, VII, and the “overlapping” mutant, whereas a significant amount of14CO2 is released in mutants from group I; (4) a substantial rate of release of14CO2 from [U-14C]aspartate; (5) in isolated mitochondria, succinate and α-glycerol phosphate stimulate O2 consumption whereas substrates which generate NADH, such as malate, do not; and (6) there is little or no rotenone-sensitive NADH oxidase activity in isolated mitochondria.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Somatic cell and molecular genetics 7 (1982), S. 219-233 
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We describe a mutant of Chinese hamster ovary cells which is resistant to elevated levels of α-methylornithine and α-difluoromethylornithine, reversible and enzymeactivated irreversible inhibitors, respectively, of the enzyme ornithine decarboxylase (ODC). The mutant cells have significantly elevated levels of enzyme activity compared to wildtype cells, but several of the physical parameters of the enzyme are completely normal: Michaelis-Menten parameter, Km, affinity for the analog, and half-life. The temporal regulation of this activity in synchronized cells is not perturbed, and the suppression of ODC activity by the addition of putrescine is still observed. Indirect experiments suggest increased concentrations of ODC mRNA in the mutant cells.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Somatic cell and molecular genetics 2 (1976), S. 331-344 
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A selection scheme based on the nutritional requirements of a previously described respiration-deficient Chinese hamster line has been used to isolate new mutants defective in oxidative energy metabolism. Three of the primary characteristics of this type of mutant are (1) a strict dependency on the continued presence of glucose for survival; (2) a drastic reduction in the rate of oxygen consumption; (3) an inhibition of Krebs cycle activity resulting in auxotrophy for asparagine and carbon dioxide. In the case of one cell line which was used (V79), up to 65% of the survivors of a selection were found to possess this phenotype after only one round of selection. By contrast, it proved much more difficult to obtain such mutants from another cell line (CCL16). A preliminary characterization of a number of these mutants is presented.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Somatic cell and molecular genetics 5 (1979), S. 225-240 
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We present here genetic experiments with a series of Chinese hamster cell mutants defective in oxidative energy metabolism. The mutations were all shown to be recessive in intraspecies hybrids. Thirty-five mutants were sorted into eight complementation groups, but one of these mutants failed to complement representatives of two distinct complementation groups. The possibility was raised that this is a cell carrying two mutations or a deletion. Because of the greatly different frequencies with which such mutants could be isolated from two different Chinese hamster cell lines, CCL16 (DON) and V79, the stability of representatives from each cell line was examined, and it was found that revertants could be obtained after treatment with mutagens, while spontaneous revertants appeared at unmeasurable or extremely low frequencies, with one exception. The mutant with a very noticeable frequency of spontaneous reversion was defective in mitochondrial protein synthesis, and the question arose whether the mutation was on the mitochondrial genome. A detailed fluctuation analysis of reversion rate and comparison with rates for other mutations was consistent with a nuclear mutation. This conclusion was supported by experiments involving fusions with cytoplasts.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 81 (1973), S. 199-216 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The isolation of a temperature sensitive cell line from the Chinese hamster line CCL39 of the American Type Culture Collection is described. At the nonpermissive temperature (39°C) the cells become attached to the surface of tissue culture dishes, but no microscopically observable colonies are formed upon prolonged incubation. Exposure to the high temperature for more than 24 hours leads to an almost complete loss in viability. A karyotypic analysis showed that this new line has lost one of the medium-sized metacentric chromosomes, although no proof is available so far to show that this loss is not simply coincidental.In nonsynchronized cultures transferred to 39°C DNA synthesis stops first, RNA synthesis shortly thereafter, while protein synthesis (turnover) continues for a longer time. After such a shift the cell number increases by less than 15% as measured with the Coulter counter.Studies with synchronized cultures give the following results: (1) one round of DNA synthesis can occur at 39°C when the cells are released from serum starvation or a hydroxyurea block, or when mitotic cells are placed at 39°C; (2) the entry of cells into metaphase of mitosis at 39°C is almost normal when the preceding time interval at 39°C is only eight hours (release of cells from G1/S boundary), but considerably reduced when the cells spend an additional 12 to 15 hours at 39°C in G1 (release from serum starvation).Infection by SV40 virus temporarily induces DNA synthesis after it has come to a stop at the nonpermissive temperature, but cells permanently transformed by SV40 still exhibit the temperature-sensitive phenotype.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 219-230 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A series of Chinese hamster cell lines were tested and found to be able to proliferate in the absence of added bicarbonate and carbondioxide if hypoxanthine and uridine were present in the medium. Conversely, cells incapable of salvaging one of these precursors, such as hypoxanthine-guanine phosphoribosyltransferase (HGPRT-) deficient cells did not multiply under these conditions.We describe another variant capable of utilizing hypoxanthine and uridine which has an absolute requirement for exogenous CO2/NaHCO3 for growth. These cells appear to be defective in the complete oxidation of pyruvate to carbondioxide, and indications are that the entry of pyruvate into the Krebs cycle is affected.
    Additional Material: 10 Ill.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 90 (1977), S. 145-160 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A temperature-sensitive mutant of Chinese hamster cells is described which has two interesting properties: (1) it is a cell cycle mutant and (2) glycoprotein synthesis appears to be affected at the non-permissive temperature (40°C). Synchronized cells shifted to 40°C in the beginning of their G1 phase do not incorporate [3H]-thymidine into DNA during the expected S-phase, but once DNA synthesis has been initiated (∼ 10 hours after termination of serum starvation) a shift to 40°C no longer leads to an arrest of DNA synthesis. Flow microfluorimetric analysis of DNA content/cell supports this conclusion and indicates that a majority of cells become arrested in the G1 phase of the cell cycle when a non-synchronized population of cells is transferred to 40°C. Apparently at all times in the cell cycle there is a drastic reduction of incorporation of labeled sugars (particularly fucose) into glycoproteins. The uptake of fucose and its conversion to GDP-fucose appears to be normal at 40°C. Chromatographic analysis indicates that all classes of glycoproteins are affected, and we do not find any evidence for partially completed oligosaccharides at 40°C. Overall protein synthesis is not reduced at the nonpermissive temperature during the time interval under consideration and the number of polysomes attached to membranes (RER) is also normal at 40°C. This suggests that the defect is at an early step in the synthesis or regulation of synthesis of glycoproteins. The mutation is a recessive mutation in hybrid cells and mutagen induced revertants can be obtained which grow normally at 40°C and in which glycoprotein synthesis at 40°C is restored to normal, wild type levels.
    Additional Material: 11 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 103 (1980), S. 169-172 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: High pressure liquid chromatography was used to determine the base, nucleoside, and nucleotide levels in wild type and a series of respiration-deficient Chinese hamster cell mutants. From this analysis the size of the total adenylate pool and the energy charge could be calculated for each cell line. We find a constant energy charge, as expected, but the adenylate pool seems to be considerably lower in the respiration-deficient mutants.
    Additional Material: 1 Ill.
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