Isolation and characterization of Staphylococcus aureus starvation-induced, stationary-phase mutants defective in survival or recovery.
Ten Staphylococcus aureus mutants, defective in the starvation-induced stationary phase of growth were isolated from two independent Tn917-LTV1 transposon insertion libraries and were designated suv as they had apparent survival defects. Seven of these mutants were defective under amino-acid-limiting conditions alone. Two mutants (suv-3 and suv-20) demonstrated lower plating efficiency when starved for glucose, phosphate or amino acids and one mutant (suv-11) had reduced plating efficiency after amino acid or glucose starvation. All of the mutants tested were as resistant to hydrogen peroxide assault as the parent, but six were more sensitive to low pH conditions. All the mutants were physically mapped on the S. aureus chromosome using PFGE. Chromosomal DNA flanking the Tn917-LTV1 insertion sites was rescued by cloning into Escherichia coli. DNA sequence analysis resulted in the identification of a number of transposon-disrupted ORFs encoding putative components such as superoxide dismutase (suv-1), haem A synthase (suv-3), a component of the SOS response (suv-9) and hypoxanthine-guanine phosphoribosyltransferase (suv-20). The Tn917-LTV1 insertion created lacZ transcriptional fusions for some of the stationary-phase loci. Expression analysis indicated that suv-4 was induced at mid-exponential phase, whereas suv-3 and suv-11 were induced at the onset of stationary phase. The possible roles of these suv components in stationary-phase survival or recovery is discussed.
Watson
Sean P
SP
Department of Molecular Biology and Biotechnology, University of Sheffield,Firth Court, Western Bank, Sheffield 510 2TN,UK.
Antonio
Martin
M
Department of Molecular Biology and Biotechnology, University of Sheffield,Firth Court, Western Bank, Sheffield 510 2TN,UK.
Foster
Simon J
SJ
Department of Molecular Biology and Biotechnology, University of Sheffield,Firth Court, Western Bank, Sheffield 510 2TN,UK.
D004251:DNA Transposable Elements / Q000235:genetics*; D016521:Electrophoresis, Gel, Pulsed-Field; D005947:Glucose / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D016254:Mutagenesis, Insertional; D010710:Phosphates / Q000378:metabolism; D020161:Physical Chromosome Mapping; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013211:Staphylococcus aureus / Q000235:genetics* / Q000254:growth & development*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
D004251:DNA Transposable Elements; D010710:Phosphates; D011993:Recombinant Fusion Proteins; D006861:Hydrogen Peroxide; D001616:beta-Galactosidase; D005947:Glucose