CARBON AND ENERGETIC UNCOUPLING ARE ASSOCIATED WITH BLOCK OF DIVISIONAT DIFFERENT STAGES OF THE CELL-CYCLE IN SEVERAL CDC MUTANTS OF SACCHAROMYCES-CEREVISIAE

Cell proliferation arrest at 37 degrees C (restrictive temperature) of the cell division cycle (cdc) mutants of Saccharomyces cerevisiae cdc 28, cdc35, cdc19, cdc21, and cdc17 was correlated with carbon and ener gy uncoupling. At 37 degrees C, cdc mutants diverted to biomass synthe sis only 3 to 4% and 8 to 24% of the fluxes of carbon consumed and ATP obtained by catabolism, respectively, compared with 48 and 34% in the wild-type strain A364A. At the permissive temperature (25 degrees C), the wild type showed similar carbon and energy coupling indexes as at 37 degrees C. However, carbon and energy coupling indexes were two- t o sevenfold higher at 25 degrees than at 37 degrees C in cdc mutants; e.g., at 25 degrees C two- to sevenfold higher amounts of carbon and A TP were directed to biomass production than at 37 degrees C. The wild- type strain exhibited a purely oxidative glucose catabolism at 37 degr ees C (RQ similar to 1.0), while the cell proliferation arrest of cdc mutants at the same temperature was characterized by fermentative meta bolism. At 37 degrees C, cdc mutants directed 50 to 60% of the carbon to ethanol production; 3 to 12% of the carbon was recovered as glycero l in cdc mutants as well as in the wild type. The proliferation arrest of the cell division cycle mutant cdc28 correlated with a significant decrease in the incorporation of radioactive precursors into DNA, RNA , and proteins. In the presence of 8-hydroxyquinoline, the wild-type s train underwent cell proliferation arrest and also exhibited metabolic uncoupling with bioenergetic and catabolic behavior similar to that o f the cdc mutants at 37 degrees C. Experimental evidence obtained with cdc19, whose defective gene product is pyruvate kinase, suggests that the primary defect of cdc mutants correlates with a metabolically, hi ghly uncoupled yeast cell. The results presented point to the existenc e of strong carbon and energy uncoupling together with cell division a rrest exhibited by cdc mutants at the restrictive temperature. The deg ree of uncoupling appears to be tuned, at least in part, by the increa se in flux of sugar catabolism through the ethanol fermentative pathwa y. (C) 1995 Academic Press, Inc.