Certain Staphylococci possess a gene called epr or lif that renders the cells resistant to lysis by glycylglycine endopeptidase. The resistance is conferred by modifying the amino acid composition of interpeptide chains in cell-wall peptidoglycan by increasing serine content and decreasing glycine content. A gene homologous to epr/lif was cloned from S. aureus RN450 genomic libraries and designated eprh. eprh was found to localize 27 bp downstream of a novel cell-wall hydrolase gene lytN, which is in the same orientation with eprh. By analogy with epr/lif, eprh is suggested to be involved in the transfer of certain amino acids, possibly serine or amino acids other than glycine, to interpeptide chains of cell-wall peptidoglycan. Unlike epr/lif, overexpression of eprh in S. aureus did not result in an increased resistance to lysostaphin. Insertional inactivation of eprh or lytN by Campbell-type integration did not affect the susceptibility of the cells to lysostaphin; either. These results suggest that eprh and lytN are not essential genes for S. aureus growth. The physiological function of eprh remains unknown. (C) 1998 Elsevier Science B.V. All rights reserved.