Inhibition of Escherichia coli protein synthesis by abortive translation of phage lambda minigenes.
Escherichia coli mutants defective in peptidyl-tRNA hydrolase activity are unable to maintain bacteriophage lambda vegetative growth. Phage mutants, named bar, overcome the host limitation to support viral growth. Multicopy expression of lambda wild-type bar regions is deleterious to hydrolase-defective cells because it provokes arrest of protein synthesis. We noticed that the bar regions include minigenes whose transcripts would contain a Shine-Dalgarno-like sequence appropriately spaced for translation from a two codon open reading frame. To investigate the mechanism of bar inhibition, we asked if transcripts of the barI region function as mRNAs in their ribosomal interactions. We found that bar-containing RNA associates with ribosomes, forms ternary initiation complexes, yields a toeprint signal, and can be removed from ribosomes by run-off translation, as authentic mRNA. Since bar-containing RNA has the properties of a messenger, we propose that its translation leads to drop-off and accumulation of peptidyl-tRNA in pth-defective cells. Starvation of the tRNA(s) sequestered in pepidyl-tRNA(s) eventually causes inhibition of protein synthesis.
Ontiveros
C
C
Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Mexico City, México.
Valadez
J G
JG
Hernández
J
J
Guarneros
G
G
D001426:Bacterial Proteins / Q000096:biosynthesis*; D010582:Bacteriophage lambda / Q000235:genetics* / Q000254:growth & development; D002265:Carboxylic Ester Hydrolases / Q000235:genetics; D002474:Cell-Free System / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D005814:Genes, Viral; D009154:Mutation; D014176:Protein Biosynthesis; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D012270:Ribosomes / Q000378:metabolism
D001426:Bacterial Proteins; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D012367:RNA, Viral; D002265:Carboxylic Ester Hydrolases; C019674:aminoacyl-tRNA hydrolase