Peptidyl transferase inhibition by the nascent leader peptide of an inducible cat gene.
The site of ribosome stalling in the leader of cat transcripts is critical to induction of downstream translation. Site-specific stalling requires translation of the first five leader codons and the presence of chloramphenicol, a sequence-independent inhibitor of ribosome elongation. We demonstrate in this report that a synthetic peptide (the 5-mer) corresponding to the N-terminal five codons of the cat-86 leader inhibits peptidyl transferase in vitro. The N-terminal 2-, 3-, and 4-mers and the reverse 5-mer (reverse amino acid sequence of the 5-mer) are virtually without effect on peptidyl transferase. A missense mutation in the cat-86 leader that abolishes induction in vivo corresponds to an amino acid replacement in the 5-mer that completely relieves peptidyl transferase inhibition. In contrast, a missense mutation that does not interfere with in vivo induction corresponds to an amino acid replacement in the 5-mer that does not significantly alter peptidyl transferase inhibition. Our results suggest that peptidyl transferase inhibition by the nascent cat-86 5-mer peptide may be the primary determinant of the site of ribosome stalling in the leader. A model based on this concept can explain the site specificity of ribosome stalling as well as the response of induction to very low levels of the antibiotic inducer.
Gu
Z
Z
Department of Biological Sciences, University of Maryland Baltimore County, Catonsville 21228.
Rogers
E J
EJ
Lovett
P S
PS
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D002701:Chloramphenicol / Q000494:pharmacology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D004352:Drug Resistance, Microbial / Q000235:genetics; D004790:Enzyme Induction; D004917:Erythromycin / Q000494:pharmacology; D007700:Kinetics; D008034:Lincomycin / Q000494:pharmacology; D008969:Molecular Sequence Data; D010458:Peptidyl Transferases / Q000037:antagonists & inhibitors* / Q000187:drug effects; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism*; D012329:RNA, Bacterial; D012270:Ribosomes / Q000378:metabolism
D021382:Protein Sorting Signals; D012329:RNA, Bacterial; D004917:Erythromycin; D002701:Chloramphenicol; D008034:Lincomycin; D015500:Chloramphenicol O-Acetyltransferase; D010458:Peptidyl Transferases