<jats:p> Methionine biosynthesis and regulation of four enzymatic steps involved in this pathway were studied in <jats:italic>Saccharomyces cerevisiae</jats:italic> , in relation to genes concerned with resistance to ethionine ( <jats:italic>eth</jats:italic> <jats:sub>1</jats:sub> and <jats:italic>eth</jats:italic> <jats:sub>2</jats:sub> ). Data presented in this paper and others favor a scheme which excludes cystathionine as an obligatory intermediate. Kinetic data are presented for homocysteine synthetase [K <jats:sub>m</jats:sub> ( <jats:italic>O</jats:italic> -acetyl- <jats:sc>l</jats:sc> -homoserine) = 7 × 10 <jats:sup>−3</jats:sup> <jats:sc>m</jats:sc> ; <jats:italic>K</jats:italic> <jats:sub>i</jats:sub> ( <jats:sc>l</jats:sc> -methionine) = 1.9 × 10 <jats:sup>−3</jats:sup> <jats:sc>m</jats:sc> ]. Enzymes catalyzing steps 3, 4, 5, and 9 were repressible by methionine. Enzyme 4 (homoserine- <jats:italic>O</jats:italic> -transacetylase) and enzyme 9 (homocysteine synthetase) were simultaneously derepressed in strains carrying the mutant allele <jats:italic>eth</jats:italic> <jats:sub>2</jats:sub> <jats:sup>r</jats:sup> . Studies on diploid strains confirmed the dominance of the eth <jats:sub>2</jats:sub> <jats:sup>s</jats:sup> allele over eth <jats:sub>2</jats:sub> <jats:sup>r</jats:sup> . Regulation of enzyme 3 (homoserine dehydrogenase) and enzyme 5 (adenosine triphosphate sulfurylase) is not modified by the allele eth <jats:sub>2</jats:sub> <jats:sup>r</jats:sup> . The other gene <jats:italic>eth</jats:italic> <jats:sub>1</jats:sub> did not appear to participate in regulation of these four steps. Gene enzyme relationship was determined for three of the four steps studied (steps 3, 4, and 9). The structural genes concerned with the steps which are under the control of <jats:italic>eth</jats:italic> <jats:sub>2</jats:sub> ( <jats:italic>met</jats:italic> <jats:sub>8</jats:sub> : enzyme 9 and <jats:italic>met</jats:italic> <jats:sub>a</jats:sub> : enzyme 4) segregate independently, and are unlinked to <jats:italic>eth</jats:italic> <jats:sub>2</jats:sub> . These results are compatible with the idea that the gene <jats:italic>eth</jats:italic> <jats:sub>2</jats:sub> is responsible for the synthesis of a pleiotropic methionine repressor and suggest the existence of at least two different methionine repressors in <jats:italic>S. cerevisiae</jats:italic> . Implications of these findings in general regulatory mechanisms have been discussed.