RpoS- and OxyR-independent induction of HPI catalase at stationary phase in Escherichia coli and identification of rpoS mutations in common laboratory strains.
A rapid spectrophotometric assay to determine the activities of HPI and HPII catalases in Escherichia coli extracts has been developed. This assay is based upon the differential heat stabilities of the two enzymes and offers significant advantages over previous methods for quantitation of their activities. Measurement of catalase activities in extracts of various mutant strains confirmed the ability of this method to accurately distinguish the two activities. Contrary to previously published results, HPI catalase activity was observed to increase at stationary phase in strains lacking the stationary-phase sigma factor sigma(s) (RpoS). This increase was independent of OxyR and also occurred in a strain lacking the HPII structural gene, katE. These results suggest a potential novel pathway for HPI induction in response to increased oxidative stress in the absence of HPII. Measurement of HPII activity in strains carrying mutations in pcm (encoding the L-isoaspartyl protein methyltransferase) and surE led to the finding that these strains also have an amber mutation in rpoS; sequencing demonstrated the presence of this mutation in several commonly used laboratory strains of E. coli, including AB1157, W1485, and JC7623.
Visick
J E
JE
Department of Chemistry and Biochemistry and Molecular Biology Institute, University of California, Los Angeles, 90095-1569, USA.
Clarke
S
S
D000135:Acid Phosphatase; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002374:Catalase / Q000378:metabolism*; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology* / Q000302:isolation & purification; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D009154:Mutation; D018384:Oxidative Stress; D010544:Peroxidases / Q000378:metabolism*; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase; D011496:Protein Methyltransferases / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics*; D012808:Sigma Factor / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D012097:Repressor Proteins; D012808:Sigma Factor; D014157:Transcription Factors; C499299:oxyR protein, E coli; C069567:sigma factor KatF protein, Bacteria; D010544:Peroxidases; C102664:hydroperoxidase II; D002374:Catalase; C076136:catalase HPI; D011496:Protein Methyltransferases; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase; D000135:Acid Phosphatase; C089634:umpG protein, E coli