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Deoxyribonucleotide biosynthesis in synchronous algae cells.

W Feller, G Schimpff-Weiland, H Follmann

    European Journal of Biochemistry
    |September 1, 1980
    PubMed
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    This study shows that deoxythymidine triphosphate (dTTP) levels regulate DNA precursor synthesis in green algae. Inhibiting thymidylate synthesis depletes dTTP, halting DNA replication and increasing reductase activity.

    Area of Science:

    • Plant molecular biology
    • Biochemistry
    • Cell cycle regulation

    Background:

    • Deoxyribonucleoside triphosphate (dNTP) pools are essential for DNA synthesis.
    • Ribonucleotide reductase is a key enzyme in dNTP biosynthesis.
    • Understanding dNTP pool regulation is crucial for comprehending DNA replication control.

    Purpose of the Study:

    • To investigate the regulation of DNA precursor biosynthesis in the green alga Scenedesmus obliquus.
    • To elucidate the role of deoxythymidine triphosphate (dTTP) in controlling ribonucleotide reductase activity.
    • To compare DNA synthesis regulation in algae with that in other eukaryotes.

    Main Methods:

    • Culturing synchronous Scenedesmus obliquus cells under controlled light/dark cycles.
    • Measuring ribonucleoside-diphosphate reductase activity and intracellular dNTP concentrations.

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  • Utilizing inhibitors of DNA synthesis, including 5-fluorodeoxyuridine, 5-fluorouracil, methotrexate, and arabinosylcytosine.

    • Main Results:

    • Peak reductase activity and dTTP concentrations coincided with DNA synthesis in the cell cycle.
    • Inhibitors of thymidylate synthesis rapidly depleted dTTP pools and inhibited DNA synthesis.
    • Inhibition of thymidylate synthesis led to increased reductase activity and dATP pools, particularly under illumination.
    • Arabinosylcytosine, a DNA replication inhibitor, did not affect reductase activity.

    Conclusions:

    • The NDP to dNDP to dNTP pathway of DNA precursor biosynthesis is active in plant cells.
    • De novo enzyme synthesis of ribonucleotide reductase in algae appears to be regulated by dTTP presence, not directly by DNA synthesis.
    • This regulatory mechanism resembles that in HeLa cells but may differ from bacterial systems.