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Nascent peptide‐dependent ribosome stalling in drug‐inducible antibiotic resistance
Author(s) -
Ramu Haripriya,
Subramanian Sai Lakshmi,
VazquezLaslop Nora,
Mankin Alexander S.
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.496.5
Subject(s) - ribosome , biology , orfs , ribosomal binding site , gene , translation (biology) , start codon , genetics , internal ribosome entry site , shine dalgarno sequence , transcription (linguistics) , microbiology and biotechnology , messenger rna , open reading frame , peptide sequence , rna , linguistics , philosophy
Ribosome‐nascent peptide interactions are involved in regulation of expression of several bacterial genes. One such example is the induction of an antibiotic resistance gene, ermC (erythromycin resistance methyltransferase). Expression of ermC is controlled by ribosome stalling at a short upstream ORF (uORF) and depends on the sequence of the nascent peptide and the presence of erythromycin. A number of other genes that confer resistance to ribosome‐targeting antibiotics are inducible, but in most cases, the underlying mechanisms are unknown. A comprehensive sequence analysis of the upstream regions of these genes showed the presence of uORFs some of which have not been identified previously. Sequences of the encoded peptides show remarkable diversity. Toe‐printing experiments in a cell‐free transcription‐translation system revealed erythromycin‐dependent ribosome stalling at a variety of these leader ORFs. Alanine scanning mutagenesis is carried out to identify the critical sequences of nascent peptides required for drug‐dependent ribosome stalling. These data are expected to provide significant insight into the molecular mechanisms of ribosome ‐ nascent peptide interactions.