The genetic basis of erythromycin resistance in cutaneous propionibacteria was determined by comparing the nucleotide sequences of the peptidyl transferase region in the 23S rRNAs from 9 susceptible and 26 resistant clinical isolates as well as 4 laboratory-selected erythromycin-resistant mutants of a susceptible strain. In 13 isolates and the 4 laboratory mutants, cross-resistance to macrolides, lincosamides, and B-type streptogramins was associated with an A --> G transition at a position cognate with Escherichia coli 23S rRNA base 2058, These strains were resistant to greater than or equal to 512 mu g of erythromycin per ml. Two other mutations were identified, an A --> G transition at base 2059 in seven strains, associated with high-level resistance to all macrolides, and a G --> A transition at base 2057 in six strains, associated with low-level resistance to erythromycin, These mutations correspond to three of four phenotypic classes previously identified by using MIC determinations.