STUDIES ON CELL METABOLISM AND CELL DIVISION : IV. COMBINED ACTION OF SUBSTITUTED PHENOLS, CYANIDE, CARBON MONOXIDE, AND OTHER RESPIRATORY INHIBITORS ON RESPIRATION AND CELL DIVISION.

The effects of 4,6-dinitro-o-cresol and 2,4,5-trichlorophenol on the respiration and cell division of fertilized eggs of Arbacia punctulata have been determined in the presence of each of a number of respiratory inhibitors. The experimental results obtained appear to afford some understanding of the mechanism of action of the substituted phenols on respiration and on cell division. 1. From the fact that the stimulated respiration is completely cyanide and carbon monoxide sensitive, it may be concluded that all of the extra oxygen uptake induced in Arbacia eggs by 4,6-dinitro-o-cresol passes through the metal containing oxidase system. All of the extra oxygen uptake also passes through oxidative steps which can be poisoned by non-stimulating phenols like 2,4-dinitrothymol and 4-nitrocarvacrol, by phenylurethane, by 5-isoamyl-5-ethyl barbituric acid, by malonic acid, or by iodoacetic acid. To abolish all respiratory stimulation by suboptimum concentrations of 4,6-dinitro-o-cresol, each of these inhibitors must be present in a concentration which reduces the normal respiration in the absence of substituted phenols by at least 20-40 per cent. 2. The degree of reduction of the stimulated respiration by a given concentration of carbon monoxide or potassium cyanide depends on the concentration of 4,6-dinitro-o-cresol or 2,4,5-trichlorophenol, being most marked in suboptimum concentrations and least marked in greater than optimum concentrations of the substituted phenol. In contrast to this result, the reduction of the stimulated respiration by a given concentration of 5-isoamyl-5-ethyl barbituric acid or malonic acid is least marked in suboptimum concentrations and most marked in greater than optimum concentrations of the substituted phenol. 3. The present experiments appear to indicate that the inhibition of cell division by substituted phenols is not attributable to a direct action of these agents on mitotic processes nor to an overstimulation of any respiratory process. The inhibition of cell division appears to be associated with the inhibition, by the substituted phenols, of some component of the cyanide sensitive respiratory system. This inhibition is of such a type as to allow the overall respiration to proceed at a rate in excess of the control value, even when division is completely suppressed. The dependence of the division mechanism on a respiratory step which is relatively hypersensitive to poisoning by the substituted phenols is comparable to the dependence of the Pasteur reaction in certain normal and tumor tissues on an oxidative step which is specifically poisoned by the substituted phenols (16). The substituted phenols have no inhibiting effect in vitro on the principal metal containing respiratory catalysts or the principal dehydrogenases; they also do not inhibit the fermentative reactions involved in the anaerobic glycolysis of fertilized Arbacia eggs. It is therefore suggested that the respiratory inhibiting and division inhibiting effects of the substituted phenols may be attributable to the action of these substances on one or more of the oxidation-reduction or phosphorylating steps which are involved in the transfer of hydrogen from the dehydrogenase systems to the specifically cyanide sensitive oxidase mechanism of the eggs. The identification of the respiratory step poisoned by the substituted phenol would constitute an interesting contribution to the chemistry of cell division and experiments to this end are now in progress.