AbstractIn the yeast Saccharomyces cerevisiae the PEL1 gene is essential for the viability of rho−/rhoo petite mutants, and its mutation in respiring cells results in a pleiotropic phenotype. Results of complementation analysis with different subclones of chromosomal DNA and re‐sequencing of the YCL4w‐YCL3w segment of chromosome III demonstrate that the coding region of the PEL1 gene corresponds to 1467 bp. The size of the PEL1 transcript in Northern blot analysis was estimated to be approximately 1·5 kb. Transcription initiation in wild‐type cells was found to occur at the position −9 relative to the ATG. The PEL1 gene was moderately expressed irrespective of the state of the mitochondrial genome and the nature of the carbon sources. Disruption of the PEL1 gene was not lethal and resulted in the same phenotype as observed with the pel1 mutant, i.e. the cells were not able to survive ethidium bromide mutagenesis, were thermosensitive for growth on glucose at 37°C and failed to grow on minimal glycerol medium. Although the Pel1 protein exhibits significant similarity to a family of phosphatidylserine synthases, the disrupted PEL1 gene was not complemented by the multicopy plasmid‐borne CHO1 gene encoding an essential yeast phosphatidylserine synthase. The Accession Number of the PEL1 gene in the EMBL data library is Z48162.