title: Residual polarity and transcription-translation coupling during recovery from chloramphenicol or fusidic acid.
abstract: Fusidic acid or chloramphenicol was used to inhibit peptide synthesis to 1% of normal in Escherichia coli B, strain AS19. After 10 min of inhibition, peptide synthesis could be quickly restored to 80% of the normal rate after washing the bacteria on a filter. However, even in the presence of adenosine 3'-5'-cyclic-monophosphoric acid to block catabolite repression, beta-galactosidase, the first enzyme of the lactose operon (lac), could only be induced to 10% of normal, and the last enzyme of the operon, galactoside acetyltransferase, even less. The first and last enzymes of the operon for tryptophan synthesis could be derepressed to about 30% of normal. The lac ribonucleic acid (RNA) induced during recovery showed a smaller than normal size distribution on sucrose gradients. The operator-proximal or -distal parts of this RNA were specifically labeled. Hybridization to phi80dlac deoxyribonucleic acid (DNA) suggested that although the distal parts of the lac RNA were barely detectable, initiation was occurring at normal rates in recovery. Either normal levels of distal messenger RNA (mRNA) are made but then rapidly degraded or the mRNA is not completed. The small amount that is made decayed abnormally slowly, probably as a result of slower transcription. Total mRNA decay was multiphasic with all components decaying slower than normal. We propose that there is a residual level of inhibition of peptide synthesis during recovery. The probability that a ribosome is blocked at any codon can be estimated from the data. The longer the message, the less likely its complete translation. We propose that the RNA polymerase can transcribe translatable mRNA for only a finite distance beyond the lead ribosome. Because ribosomes can load at the start of each message in a polycistronic mRNA, the probability that a distal message will be synthesized and translated is a function of the number of more proximal messages and the distances between their ribosome-loading sites.
mesh_terms: D000123:Acetyltransferases / Q000096:biosynthesis; D000878:Anthranilate Synthase / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000096:biosynthesis; D002250:Carbon Radioisotopes; D002499:Centrifugation, Density Gradient; D002701:Chloramphenicol / Q000494:pharmacology*; D000242:Cyclic AMP / Q000494:pharmacology; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism*; D005672:Fusidic Acid / Q000494:pharmacology*; D005696:Galactosidases / Q000096:biosynthesis; D007785:Lactose / Q000378:metabolism; D007930:Leucine / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D009876:Operon; D010452:Peptide Biosynthesis; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis / Q000378:metabolism; D014158:Transcription, Genetic; D014316:Tritium; D014364:Tryptophan / Q000096:biosynthesis; D014367:Tryptophan Synthase / Q000096:biosynthesis; D014498:Uracil / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D002250:Carbon Radioisotopes; D012329:RNA, Bacterial; D012333:RNA, Messenger; D014316:Tritium; D014498:Uracil; D005672:Fusidic Acid; D002701:Chloramphenicol; D014364:Tryptophan; D000242:Cyclic AMP; D000123:Acetyltransferases; D005696:Galactosidases; D000878:Anthranilate Synthase; D014367:Tryptophan Synthase; D007930:Leucine; D007785:Lactose