Uptake of Uracil by Synchronous Chlorella fusca

Uptake of radioactive uracil by light-dark synchronized Cblorella fusca Shihira and Krauss was studied. For the characterization of the uptake system autospores were used and the following results obtained. Autospores kept in the dark accumulated uracil against a concentration gradient in a process having an observed activation energy of 10 keal/mol in the 10–40°C interval. Addition of glucose to the reaction suspension did not affect the uptake, but, 100 γM dinitrophenol inhibited the process by 90%. Abrupt changes in rate were found upon changing the conditions from light to dark and vice versa, and the rates measured in light were about 2.5 times larger than those found in the dark. Initial rates measured in the dark followed saturation kineties with half maximal rate found at 0.25 γM uracil, and with an apparent maximal rate of 1.7.10^-10 mol/10 min . 10⁷ cells. The effect of 14 pyrimidines on uptake was tested, and it was found that uracils which were substituted in the 5′ or 5′+ 6′ positions were strongly inhibitory. Of these, thymine and dihydrouracil were tested and shown to inhibit uracil uptake competitively. Initial uptake rates, measured in the dark with 1.0 γM uracil, were recorded at intervals during the 24 h synchronous cycle. The uptake rate per ml culture was constant during the first 9 h, thereafter increasing to reach a peak value at 14 h. This peak was followed by a strong increase from 18 h onwards, this increase being concomitant with the sporulation process, and closely followed its time course.