Open AccessDeoxyribonucleotide Biosynthesis in Synchronous Algae Cells
Author(s) -
FELLER Wolfgang,
SCHIMPFFWEILAND Gabriele,
FOLLMANN Hartmut
Publication year - 1980
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1980.tb04843.x
Subject(s) - ribonucleotide reductase , dna synthesis , cycloheximide , biochemistry , ribonucleotide , biology , deoxyribonucleotide , de novo synthesis , biosynthesis , ribonucleoside , nucleotide , dna , microbiology and biotechnology , enzyme , protein biosynthesis , rna , protein subunit , gene
Synchronous cells of the green alga, Scenedesmus obliquus , cultured in a 14‐h/10‐h light/dark regime, contain a peak of ribonucleoside‐diphosphate reductase activity and maximum deoxyribonucleoside 5′‐triphosphate concentrations at the 12th hour of the cell cycle, coinciding with DNA synthesis and preceding the formation of eight daughter cells. The intracellular dTTP pool reaches 4.5 pmol and the other pools 2–3 pmol/10 6 cells. Algal reductase activity is sensitive to cycloheximide, but not to lincomycin. These correlations demonstrate the functioning of the NDP → dNDP → dNTP pathway of DNA precursor biosynthesis in plant cells. In the presence of 20 μg 5‐fluorodeoxyuridine/ml, an inhibitor of thymidylate synthesis, the dTTP pool is rapidly depleted and DNA synthesis ceases. 5‐Fluorouracil and methotrexate produce similar effects. At the same time the ribonucleotide reductase activity and also the dATP pool are greatly increased, especially when fluorodeoxyuridine treatment is combined with continued illumination of the algae. In contrast, arabinosylcytosine, an inhibitor of DNA replication, has no effect on ribonucleotide reduction. The control of de novo enzyme synthesis in the eucaryotic algae therefore appears to depend on the presence of dTTP (or a related nucleotide), but not directly coupled to DNA synthesis. This interdependence resembles the situation observed in HeLa cells, while it may differ in detail from control mechanisms of ribonucleotide reductase studied in bacteria.