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  • Genetics  (1)
  • polyamines  (1)
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  • 1
    ISSN: 1573-5028
    Keywords: Solanum tuberosum ; yeast ; polyamines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract S-adenosylmethionine decarboxylase (SAMDC) is involved in the biosynthesis of the polyamines, spermidine and spermine. Recently, we reported the isolation of a putative cDNA clone of the SAMDC clone of potato (Plant Mol Biol 20; 641–651). In order to confirm that the potato genes does encode SAMDC, a complementation experiment with a yeast strain that possesses a null mutation in the SAMDC gene was performed. The yeast strain contains a deletion-insertion mutation in the SAMDC gene and has an absolute requirement for the addition of exogenous spermidine for growth. When the full-length potato cDNA was expressed in the mutant yeast strain there was no longer a requirement for exogenous spermidine. Immunoblotting experiments suggest that the potato SAMDC gene product has an apparent molecular mass of 39 kDa. Expression of the SAMDC gene was high in the young and actively dividing tissues and low in the mature and non-dividing tissues of both vegetative and reproductive organs. Additionally, isolation and characterisation of the corresponding genomic clone is reported. The gene has one intron in its 5′-untranslated sequence but otherwise the transcribed portion is identical to the cDNA clone.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0749-503X
    Keywords: Cell cycle ; toxin ; K. lactis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Kluyveromyces lactis toxin is a heterotrimeric protein which irreversible arrests proliferation of sensitive Saccharomyces cerevisiae cells in the G1 phase of the cell cycle. By expressing the γ subunit of the toxin in sensitive yeast cells from a conditional promoter, it was previously demonstrated that it alone is required for inhibition (Tokunaga et al. (1989). Nucleic Acids Res. 17, 3435-3446). Here we show that, like native exogenous toxin, intracellular γ subunit expression promoters a striking arrest of sensitive cells in G1. However, unlike the G1 arrest caused by native toxin, that induced by the γ subunit alone does not result in reduced cellular viability and is fully and rapidly reversible, suggesting that the G1 arrest and the irreversibility of action may reflect different aspects of the toxin's interaction with sensitive cells. We have selected a large number of S. cerevisiae mutants which are highly resistant to the toxin in order to study its mode of action in more detail. Complementation analysis demonstrated that all but one of the mutants were recessive and these defined four separate genes. Members of two complementation groups concurrently acquired resistance to intracellular γ subunit expression, suggesting that they contain a modified toxin target site. The other two genes appear to be required for entry of the γ subunit into the sensitive cell since these mutants, while refractory to exogenous toxin, were fully sensitive to intracellular γ subunit expression.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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