Anti-peptidyl transferase leader peptides of attenuation-regulated chloramphenicol-resistance genes.
The chloramphenicol (Cm)-inducible cmlA gene of Tn1696 specifies nonenzymatic resistance to Cm and is regulated by attenuation. The first eight codons of the leader specify a peptide that inhibits peptidyl transferase in vitro. Functionally similar, but less inhibitory, peptides are encoded by the leaders of Cm-inducible cat genes. However, the cat and cmlA coding sequences are unrelated and specify proteins of unrelated function. The inhibition of peptidyl transferase by the leader peptides is additive with that of Cm. Erythromycin competes with the inhibitory action of the peptides, and erythromycin and the peptides footprint to overlapping sites at the peptidyl transferase center of 23S rRNA. It is proposed that translation of the cmlA and cat leaders transiently pauses upon synthesis of the inhibitor peptides. The predicted site of pausing is identical to the leader site where long-term occupancy by a ribosome (ribosome stalling) will activate downstream gene expression. We therefore propose the inducer, Cm, converts a peptide-paused ribosome to the stalled state. We discuss the idea that cooperativity between leader peptide and inducer is necessary for ribosome stalling and may link the activation of a specific drug-resistance gene with a particular antibiotic.
Gu
Z
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Department of Biological Sciences, University of Maryland Baltimore County, Catonsville 21228.
Harrod
R
R
Rogers
E J
EJ
Lovett
P S
PS
D000595:Amino Acid Sequence; D001412:Bacillus subtilis; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics*; D002702:Chloramphenicol Resistance; D004640:Emetine / Q000494:pharmacology; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects*; D010458:Peptidyl Transferases / Q000037:antagonists & inhibitors*; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*; D013824:Thermus
D012329:RNA, Bacterial; D012333:RNA, Messenger; D012338:RNA, Ribosomal, 23S; D015500:Chloramphenicol O-Acetyltransferase; D010458:Peptidyl Transferases; D004640:Emetine