A short DNA sequence from lambda phage inhibits protein synthesis in Escherichia coli rap.
The Escherichia coli rap mutant inhibits vegetative growth of bacteriophage lambda. Phage mutations termed bar, which overcome the rap defect, have been mapped to three genetic loci in the pL operon. Plasmids with a lambda wild-type bar DNA segment cloned downstream from an active promoter cannot be maintained in rap mutant bacteria. The viability of a rap mutant strain decreases rapidly after induction of transcription through bar regions present on plasmids. Under these (restrictive) conditions the expression of plasmid-encoded beta-lactamase and plasmid DNA replication are arrested, but plasmid RNA synthesis continues for several hours. Analysis of protein extracts from E. coli rap cells containing bar plasmids revealed that both plasmid and bacterial protein synthesis are inhibited under restrictive conditions. In addition, unlike other RNAs tested, the chemical half-life of bar RNA increases 3.5-fold relative to the half-life of bar RNA under permissive conditions. We propose that transcription through the bar region, or the accumulation of bar RNA, results in an irreversible defect in cellular mRNA translation. This defect eventually kills the rap cells, and thus prevents bar plasmid maintenance.
Pérez-Morga
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Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del 1PN, Mexico D.F.
Guarneros
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D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D010582:Bacteriophage lambda / Q000235:genetics*; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D009154:Mutation; D010957:Plasmids; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004279:DNA, Viral