Expulsion of uracil and thymine from the yeast Saccharomyces cerevisiae: contrasting responses to changes in the proton electrochemical gradient.

Abstract

The outflow of uracil from the yeast Saccharomyces cerevisiae is known to be relatively fast in certain circumstances, to be retarded by proton conductors and to occur in strains lacking a uracil proton symport. In the present work, it was shown that uracil exit from washed yeast cells is an active process, creating a uracil gradient of the order of -80 mV relative to the surrounding medium. Glucose accelerated uracil exit, while retarding its entry. DNP or sodium azide each lowered the gradient to about -30 mV, simultaneously increasing the rate of uracil entry. They also lowered cellular ATP content. Manipulation of the external ionic conditions governing delta mu H+ at the plasma membrane had no detectable effect on uracil transport in yeast preparations thoroughly depleted of ATP. It was concluded that uracil exit is probably not driven by the proton gradient but may utilize ATP directly. It is known that thymine is not normally absorbed by yeast. However, thymine expulsion was here observed during deamination of the substrate 5-methylcytosine in the presence of glucose. In the absence of glucose, or following ATP depletion, thymine uptake from the medium only occurred when delta mu H+ was dissipated, either by DNP or azide, or by manipulation of the external ionic environment. The yeast expelled absorbed thymine when delta mu H+ was restored to the physiological range. The properties of the system corresponded to those of an H+/thymine antiport that is distinct from the mechanism expelling uracil.