The expression of gene CPA1, encoding the glutaminase subunit of the arginine pathway carbamoyl-phosphate synthetase, is repressed by arginine at a posttranscriptional level. The 5' region of CPA1 mRNA contains a 25 codon upstream open reading frame. The importance of this feature for the repression of CPA1 expression has been analyzed by oligonucleotide-directed mutagenesis and by sequencing of constitutive cis-dominant mutations obtained in vivo. The results show that the leader peptide, the product of the upstream open reading frame, plays an essential, negative role in the specific repression of CPA1 by arginine. A model of translational regulation of CPA1 is proposed that takes into account the cis-dominance of the mutations affecting the leader peptide.