Interfering with different steps of protein synthesis explored by transcriptional profiling of Escherichia coli K-12.
Escherichia coli responses to four inhibitors that interfere with translation were monitored at the transcriptional level. A DNA microarray method provided a comprehensive view of changes in mRNA levels after exposure to these agents. Real-time reverse transcriptase PCRanalysis served to verify observations made with microarrays, and a chromosomal grpE::lux operon fusion was employed to specifically monitor the heat shock response. 4-Azaleucine, a competitive inhibitor of leucyl-tRNA synthetase, surprisingly triggered the heat shock response. Administration of mupirocin, an inhibitor of isoleucyl-tRNA synthetase activity, resulted in changes reminiscent of the stringent response. Treatment with kasugamycin and puromycin (targeting ribosomal subunit association as well as its peptidyl-transferase activity) caused accumulation of mRNAs from ribosomal protein operons. Abundant biosynthetic transcripts were often significantly diminished after treatment with any of these agents. Exposure of a relA strain to mupirocin resulted in accumulation of ribosomal protein operon transcripts. However, the relA strain's response to the other inhibitors was quite similar to that of the wild-type strain.
Sabina
Jeffrey
J
Central Research and Development, DuPont Company, Wilmington, Delaware 19880-0173, USA
Dover
Nir
N
Templeton
Lori J
LJ
Smulski
Dana R
DR
Söll
Dieter
D
LaRossa
Robert A
RA
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D007930:Leucine / Q000031:analogs & derivatives* / Q000494:pharmacology; D016712:Mupirocin / Q000494:pharmacology; D020411:Oligonucleotide Array Sequence Analysis; D014176:Protein Biosynthesis / Q000187:drug effects*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011691:Puromycin / Q000494:pharmacology; D014158:Transcription, Genetic
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D011500:Protein Synthesis Inhibitors; C000111:azaleucine; D011691:Puromycin; D016712:Mupirocin; D007930:Leucine; C100305:kasugamycin