Attenuation control of the Escherichia coli phenylalanyl-tRNA synthetase operon.
The pheST operon codes for the two subunits of the essential enzyme phenylalanyl-tRNA synthetase. The nucleotide sequence of the regulatory regions of the operon, in vitro transcription data and in vivo experiments indicate that the operon is controlled by attenuation in a way similar to many amino acid biosynthetic operons. In this work the control of the pheST operon was studied in vivo by measuring the effect of deletions in the regulatory regions on downstream expression. The presence of a strong promoter followed by an approximately 90% efficient terminator in front of the structural parts of the operon is demonstrated. An open reading frame coding for a 14 amino acid long leader peptide containing five phenylalanine residues is located between the promoter and the terminator. The presence of the transcription terminator is shown to be essential to the operon's regulation. The localization of the promoter and the terminator agrees with the results of previous in vitro experiments. It is also shown that about 30% of the transcripts covering the pheST operon come from the upstream gene, rplT, which codes for the ribosomal protein L20. Although cotranscription exists between rplT and pheST, these genes are not systematically coregulated since reducing the translation of rplT about tenfold, does not change pheST expression. The pheST operon is also shown to be derepressed by a cellular excess of phenylalanyl-tRNA synthetase. This derepression is shown to be due to the pheST attenuator.
Springer
M
M
Mayaux
J F
JF
Fayat
G
G
Plumbridge
J A
JA
Graffe
M
M
Blanquet
S
S
Grunberg-Manago
M
M
D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics*; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D004279:DNA, Viral; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005786:Gene Expression Regulation; D005814:Genes, Viral; D009876:Operon; D010652:Phenylalanine-tRNA Ligase / Q000235:genetics*; D010957:Plasmids; D014176:Protein Biosynthesis; D014158:Transcription, Genetic
D004279:DNA, Viral; D000604:Amino Acyl-tRNA Synthetases; D010652:Phenylalanine-tRNA Ligase