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pubmed:abstractText |
In vivo studies have shown that, in the absence of homoserine-O-transacetylase activity (locus met(2)), the C(4)-carbon moiety of ethionine is utilized (provided the ethionine resistance gene eth-2r is present) by methionine auxotrophs, except for met(8) mutants (homocysteine synthetase-deficient). Concomitant utilization of sulfur and methyl group from methylmercaptan or S-methylcysteine has been demonstrated. In the absence of added methylated intermediates, the methyl group of methionine formed from ethionine is derived from serine. In vitro studies with crude extracts of Saccharomyces cerevisiae have demonstrated that this synthesis of methionine occurs by the following reactions: CH(3)-SH + ethionine right harpoon over left harpoon methionine + C(2)H(5)SH and S-methylcysteine + ethionine right harpoon over left harpoon methionine + S-ethylcysteine. In the forward direction, the second product of the second reaction was shown to be S-ethylcysteine; this reaction has also been found reversible, leading to ethionine formation. Genetic and kinetic data have shown that homocysteine synthetase catalyzes these two reactions, at 0.3% of the rate it catalyzes direct homocysteine synthesis: O-Ac-homoserine + Na(2)S --> homocysteine + acetate. The three reactions are lost together in a met(8) mutant and are recovered to the same extent in spontaneous prototrophic revertants from this strain. Methionine-mediated regulation of enzyme synthesis affects the three activities and is modified to the same extent by the presence of the recessive allele (eth-2r) of the regulatory gene eth-2. Affinities of the enzyme for substrates of both types of reactions are of the same order of magnitude. Moreover, ethionine, the substrate of the second reaction, inhibits the third reaction, whereas O-acetyl-homoserine, the substrate of the third reaction, inhibits the second reaction. An enzymatic cleavage of S-methylcysteine, leading to methylmercaptan production, has been shown to occur in crude yeast extracts. It is concluded that the enzyme homocysteine synthetase participates in the two alternate pathways leading to methionine biosynthesis in S. cerevisiae, one involving O-acetyl-homoserine and H(2)S, the other involving the 4-carbon chain of ethionine and a mercaptyl donor. Participation of the two types of reactions catalyzed by homocysteine synthetase, in in vivo methionine synthesis, has been shown to occur in a met(2) partial revertant.
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