Macrolides, lincosamides, and streptogramin B (MLSB) are structurally distinct antibiotics that are clinically successful in treatment of bacterial infections. Their efficacy has decreased due to the rapid emergence of resistant bacterial strains. Cross‐resistance to MLSB antibiotics often arises through the production of erythromycin resistance N‐6‐methyltransferase (Erm). The enzyme modifies the common MLSB binding site within the ribosomal exit tunnel and reduces the drug binding affinity to the ribosome. Previous model based on the ermCL‐ermC system suggests that the synthesis of Erm is induced through the antibiotic‐stalled ribosome that translates the co‐transcribed upstream leader sequence. Ribosome stalling induces a structural rearrangement of erm transcript and exposes the otherwise masked erm Shine‐Dalgarno (SD) sequence, subsequently leading to translational initiation. Curiously, many resistant clinical isolates carry mutated versions of leader sequence, including the ribosome‐stalling null mutants. The seemingly paradoxical role of leader peptide‐dependent ribosome stalling in MLSB resistance remains poorly understood.We used the ermBLEF‐ermB operon from the human pathogen Staphylococcus aureus as a model to determine how the expression of ermB, unlike other homologous erm genes, is inducible by a broader spectrum of MLSB antibiotics. Our data show that disrupting the translation of ErmBL abolishes ribosome stalling but results in high basal levels of ErmB that is well correlated with elevated ribosome methylation and antibiotic resistance, suggesting that a ribosome stalling‐independent determinant is required for ermB expression. Subsequently, we found that both MLSB antibiotic treatment and abrogating ErmBL translation upregulate the basal ermB mRNA levels and prolong the ermB mRNA half‐life. Furthermore, antibiotic exposure reduces mRNA cleavage at a site immediately downstream of the ermB SD sequence. The data confirmed that the expression of ermB could be upregulated by antibiotics via stabilization and accumulation of its mRNA. Finally, genome‐wide transcriptome analysis showed that the stabilizing effect of antibiotics on mRNA is not limited to ermBL‐ermB; antibiotics representing a ribosome stalling inducer and a non‐inducer co‐regulate similar sets of genes and increase the half‐lives of the majority of analyzed transcripts.Collectively, our data suggest a model by which the upregulation of ErmB is achieved through disrupted translation of the leader peptide, which normally acts as a break to fine‐tune the cellular ErmB levels. Tight regulation of ErmB is necessary because hypermethylation of ribosome may impose a fitness cost on the bacterium.Support or Funding InformationThis work is supported by the NIH, the Pew Charitable Trust (Pew Scholars in the Biomedical Sciences), the Edward Mallinckrodt Jr. Foundation, and SLU President's Research Fund.