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Regulation of polyamine synthesis in relation to putrescine and spermidine pools in Neurospora crassa.


Polyamine pools were measured under various conditions of high and low concentrations of cytosolic ornithine with the wild-type and mutant strains of Neurospora crassa. In minimal medium, the wild-type strain has 1 to 2 nmol of putrescine and approximately 14 nmol of spermidine per mg (dry weight); no spermine is found in N. crassa. Exogenous ornithine was found to cause a rapid, but quickly damped, increase in the rate of polyamine synthesis. This effect was greater in a mutant (ota) unable to catabolize ornithine. No turnover of polyamines was detected during exponential growth. Exogenous spermidine was not taken up efficiently by N. crassa; thus, the compound could not be used directly in studies of regulation. However, by nutritional manipulation of a mutant strain, aga, lacking arginase, cultures were starved for ornithine and thus ultimately for putrescine and spermidine. During ornithine starvation, the remaining putrescine pool was not converted to spermidine. The pattern of polyamine synthesis after restoration of ornithine to the polyamine-deprived aga strain indicated that, in vivo, spermidine regulates polyamine synthesis at the ornithine decarboxylase reaction. The results suggest that the regulatory process is a form of negative control which becomes highly effective when spermidine exceeds its normal level. The possible relationship between the regulation of polyamine synthesis and the ratio of free to bound spermidine is discussed.

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