UC Irvine

In Neurospora crassa mycelia, the amounts of the main polyamines, putrescine and spermidine, are approximately 0.8 and 18 nmol/mg, dry weight. We wished to know what determines these pool sizes. In the growth medium, externally added polyamines enter cells largely by a nonsaturable, diffusional system. In a mutant unable to synthesize polyamines, internal and external spermidine appear to equilibrate across the cell membrane during growth. However, this was true only after an intracellular "sink," with a capacity equal to the amount of spermidine found in wild-type cells, had been saturated. We speculate that internal anionic binding sites, detectable in permeabilized cells, sequester virtually all of the spermidine normally found in exponentially growing N. crassa. Further evidence for this view was that in mature, stationary cultures, excess spermidine is excreted. Putrescine is also excreted if its concentration in the cell is abnormally high. The control of pool size by intracellular binding and excretion may be an advantage in this pathway, because feedback inhibition does not prevail, enzyme regulation is by comparison slow, and excessive polyamines are toxic. © 1989.