Role of the hprT–ftsH locus in Staphylococcus aureus

Microbiology ◽  
2004 ◽  
Vol 150 (2) ◽  
pp. 373-381 ◽  
Author(s):  
James K. Lithgow ◽  
Eileen Ingham ◽  
Simon J. Foster
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Methyl Viologen ◽  
Potassium Tellurite ◽  
Mutagenesis Screen ◽  
Acid Methyl ◽  
Starvation Survival ◽  
Membrane Bound ◽  
Hypoxanthine Guanine Phosphoribosyltransferase

The roles of two adjacent genes in the Staphylococcus aureus chromosome with functions in starvation survival and the response to stressful conditions have been characterized. One of these, hprT, encoding a hypoxanthine–guanine phosphoribosyltransferase homologue, was initially identified in a transposon mutagenesis screen. Mutation of hprT affects starvation survival in amino-acid-limiting conditions and the ability of S. aureus to grow in high-salt concentrations. Downstream of hprT is ftsH, which encodes a membrane-bound, ATP- and Zn2+-dependent ‘AAA’-type protease. Mutation of ftsH in S. aureus leads to pleiotropic defects including slower growth, sensitivity to salt, acid, methyl viologen and potassium tellurite stresses, and reduced survival in amino-acid- or phosphate-limiting conditions. Both hprT–lacZ and ftsH–lacZ gene fusions are expressed maximally in the post-exponential phase of growth. Although secretion of exoproteins is not affected, an ftsH mutant is attenuated in a murine skin lesion model of pathogenicity.

scholarly journals Dual Role of the Oligopeptide Permease Opp3 during Growth of Staphylococcus aureus in Milk

2009 ◽  
Vol 75 (10) ◽  
pp. 3355-3357 ◽  
Author(s):  
Elise Borez�e-Durant ◽  
Aurelia Hiron ◽  
Jean-Christophe Piard ◽  
Vincent Juillard
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Nitrogen Nutrition ◽  
Dual Role ◽  
Diauxic Growth ◽  
Extracellular Proteases ◽  
Expression Levels ◽  
Oligopeptide Permease

ABSTRACT Staphylococcus aureus RN6390 presents a diauxic growth in milk, due to amino acid limitation. Inactivation of the oligopeptide permease Opp3 (dedicated to the nitrogen nutrition of the strain) not only affects the growth of the strain but also results in reduced expression levels of three major extracellular proteases.

scholarly journals The Staphylococcus aureus Alternative Sigma Factor ςB Controls the Environmental Stress Response but Not Starvation Survival or Pathogenicity in a Mouse Abscess Model

1998 ◽  
Vol 180 (23) ◽  
pp. 6082-6089 ◽  
Author(s):  
Pan F. Chan ◽  
Simon J. Foster ◽  
Eileen Ingham ◽  
Mark O. Clements
Keyword(s):  
Staphylococcus Aureus ◽  
Stress Response ◽  
Environmental Stress ◽  
Sigma Factor ◽  
Osmotic Shock ◽  
Alternative Sigma Factor ◽  
Dependent Manner ◽  
Virulence Determinant ◽  
Starvation Survival

ABSTRACT The role of ςB, an alternative sigma factor ofStaphylococcus aureus, has been characterized in response to environmental stress, starvation-survival and recovery, and pathogenicity. ςB was mainly expressed during the stationary phase of growth and was repressed by 1 M sodium chloride. AsigB insertionally inactivated mutant was created. In stress resistance studies, ςB was shown to be involved in recovery from heat shock at 54°C and in acid and hydrogen peroxide resistance but not in resistance to ethanol or osmotic shock. Interestingly, S. aureus acquired increased acid resistance when preincubated at a sublethal pH 4 prior to exposure to a lethal pH 2. This acid-adaptive response resulting in tolerance was mediated viasigB. However, ςB was not vital for the starvation-survival or recovery mechanisms. ςB does not have a major role in the expression of the global regulator of virulence determinant biosynthesis, staphylococcal accessory regulator (sarA), the production of a number of representative virulence factors, and pathogenicity in a mouse subcutaneous abscess model. However, SarA upregulates sigB expression in a growth-phase-dependent manner. Thus, ςB expression is linked to the processes controlling virulence determinant production. The role of ςB as a major regulator of the stress response, but not of starvation-survival, is discussed.

scholarly journals Characterization of the Major Superoxide Dismutase of Staphylococcus aureus and Its Role in Starvation Survival, Stress Resistance, and Pathogenicity

1999 ◽  
Vol 181 (13) ◽  
pp. 3898-3903 ◽  
Author(s):  
Mark O. Clements ◽  
Sean P. Watson ◽  
Simon J. Foster
Keyword(s):  
Staphylococcus Aureus ◽  
Methyl Viologen ◽  
Acid Stress ◽  
Superoxide Dismutases ◽  
Phase Growth ◽  
Sod Activity ◽  
Transposon Insertion ◽  
Starvation Survival

ABSTRACT A Staphylococcus aureus mutant (SPW1) which is unable to survive long-term starvation was shown to have a transposon insertion within a gene homologous to the sodA family of manganese-dependent superoxide dismutases (SOD). Whole-cell lysates of the parental 8325-4 strain demonstrated three zones of SOD activity by nondenaturing gel electrophoresis. The activities of two of these zones were dependent on manganese for activity and were absent in SPW1. The levels of SOD activity and sodA expression were growth-phase dependent, occurring most during postexponential phase. This response was also dependent on the level of aeration of the culture, with highest activity and expression occurring only under high aeration. Expression of sodA and, consequently, SOD activity could be induced by methyl viologen but only during the transition from exponential- to postexponential-phase growth. SPW1 was less able to survive amino acid limitation and acid stress but showed no alteration in pathogenicity in a mouse abscess model of infection compared to the parental strain 8325-4.

scholarly journals Characterization of the Starvation-Survival Response of Staphylococcus aureus

1998 ◽  
Vol 180 (7) ◽  
pp. 1750-1758 ◽  
Author(s):  
Sean P. Watson ◽  
Mark O. Clements ◽  
Simon J. Foster
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Nutrient Limitation ◽  
Protein Biosynthesis ◽  
Phosphate Limitation ◽  
Term Survival ◽  
Acid Shock ◽  
Nonculturable State ◽  
Starvation Survival ◽  
Survival Potential

ABSTRACT The starvation-survival response of Staphylococcus aureus as a result of glucose, amino acid, phosphate, or multiple-nutrient limitation was investigated. Glucose and multiple-nutrient limitation resulted in the loss of viability of about 99 to 99.9% of the population within 2 days. The remaining surviving cells developed increased survival potential, remaining viable for months. Amino acid or phosphate limitation did not lead to the development of a stable starvation-survival state, and cells became nonculturable within 7 days. For multiple-nutrient limitation, the development of the starvation-survival state was cell density dependent. Starvation survival was associated with a decrease in cell size and increase in resistance to acid shock and oxidative stress. There was no evidence for the formation of a viable but nonculturable state during starvation as demonstrated by flow cytometry. Long-term survival of cells was dependent on cell wall and protein biosynthesis. Analysis of [35S]methionine incorporation and labelled proteins demonstrated that differential protein synthesis occurred deep into starvation.

scholarly journals Role of a Cysteine Synthase in Staphylococcus aureus

2004 ◽  
Vol 186 (6) ◽  
pp. 1579-1590 ◽  
Author(s):  
James K. Lithgow ◽  
Emma J. Hayhurst ◽  
Gerald Cohen ◽  
Yair Aharonowitz ◽  
Simon J. Foster
Keyword(s):  
Staphylococcus Aureus ◽  
Sole Source ◽  
Knockout Mutant ◽  
Cysteine Synthase ◽  
Potassium Tellurite ◽  
E Coli ◽  
Cell Extracts ◽  
Increased Sensitivity ◽  
Thiol Content

ABSTRACT The gram-positive human pathogen Staphylococcus aureus is often isolated with media containing potassium tellurite, to which it has a higher level of resistance than Escherichia coli. The S. aureus cysM gene was isolated in a screen for genes that would increase the level of tellurite resistance of E. coli DH5α. The protein encoded by S. aureus cysM is sequentially and functionally homologous to the O-acetylserine (thiol)-lyase B family of cysteine synthase proteins. An S. aureus cysM knockout mutant grows poorly in cysteine-limiting conditions, and analysis of the thiol content in cell extracts showed that the cysM mutant produced significantly less cysteine than wild-type S. aureus SH1000. S. aureus SH1000 cannot use sulfate, sulfite, or sulfonates as the source of sulfur in cysteine biosynthesis, which is explained by the absence of genes required for the uptake and reduction of these compounds in the S. aureus genome. S. aureus SH1000, however, can utilize thiosulfate, sulfide, or glutathione as the sole source of sulfur. Mutation of cysM caused increased sensitivity of S. aureus to tellurite, hydrogen peroxide, acid, and diamide and also significantly reduced the ability of S. aureus to recover from starvation in amino acid- or phosphate-limiting conditions, indicating a role for cysteine in the S. aureus stress response and survival mechanisms.

scholarly journals A Membrane-Bound Flavocytochromec-Sulfide Dehydrogenase from the Purple Phototrophic Sulfur Bacterium Ectothiorhodospira vacuolata

2000 ◽  
Vol 182 (11) ◽  
pp. 3097-3103 ◽  
Author(s):  
Vesna Kostanjevecki ◽  
Ann Brigé ◽  
Terrance E. Meyer ◽  
Michael A. Cusanovich ◽  
Yves Guisez ◽  
...  
Keyword(s):  
Amino Acid ◽  
Signal Sequence ◽  
Sulfide Oxidation ◽  
Three Dimensional ◽  
Chromatium Vinosum ◽  
Dimensional Structure ◽  
Northern Hybridization ◽  
Amino Acid Residues ◽  
Membrane Bound

ABSTRACT The amino acid sequence of Ectothiorhodospira vacuolatacytochrome c-552, isolated from membranes withn-butanol, shows that it is a protein of 77 amino acid residues with a molecular mass of 9,041 Da. It is closely related to the cytochrome subunit of Chlorobium limicola f. sp.thiosulfatophilum flavocytochrome c-sulfide dehydrogenase (FCSD), having 49% identity. These data allowed isolation of a 5.5-kb subgenomic clone which contains the cytochrome gene and an adjacent flavoprotein gene as in other species which have an FCSD. The cytochrome subunit has a signal peptide with a normal cleavage site, but the flavoprotein subunit has a signal sequence which suggests that the mature protein has an N-terminal cysteine, characteristic of a diacyl glycerol-modified lipoprotein. The membrane localization of FCSD was confirmed by Western blotting with antibodies raised against Chromatium vinosum FCSD. When aligned according to the three-dimensional structure of ChromatiumFCSD, all but one of the side chains near the flavin are conserved. These include the Cys 42 flavin adenine dinucleotide binding site; the Cys 161-Cys 337 disulfide; Glu 167, which modulates the reactivity with sulfite; and aromatic residues which may function as charge transfer acceptors from the flavin-sulfite adduct (C. vinosumnumbering). The genetic context of FCSD is different from that in other species in that flanking genes are not conserved. The transcript is only large enough to encode the two FCSD subunits. Furthermore, Northern hybridization showed that the production of E. vacuolata FCSD mRNA is regulated by sulfide. All cultures that contained sulfide in the medium had elevated levels of FCSD RNA compared with cells grown on organics (acetate, malate, or succinate) or thiosulfate alone, consistent with the role of FCSD in sulfide oxidation.

scholarly journals Role of the (p)ppGpp Synthase RSH, a RelA/SpoT Homolog, in Stringent Response and Virulence of Staphylococcus aureus

2010 ◽  
Vol 78 (5) ◽  
pp. 1873-1883 ◽  
Author(s):  
Tobias Geiger ◽  
Christiane Goerke ◽  
Michaela Fritz ◽  
Tina Schäfer ◽  
Knut Ohlsen ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Acid Metabolism ◽  
Stringent Response ◽  
Essential Amino Acids ◽  
Transcriptional Analysis ◽  
Wild Type ◽  
Stringent Control ◽  
Nutrient Limitations

ABSTRACT In most bacteria, nutrient limitations provoke the stringent control through the rapid synthesis of the alarmones pppGpp and ppGpp. Little is known about the stringent control in the human pathogen Staphylococcus aureus, partly due to the essentiality of the major (p)ppGpp synthase/hydrolase enzyme RSH (RelA/SpoT homolog). Here, we show that mutants defective only in the synthase domain of RSH (rsh syn) are not impaired in growth under nutrient-rich conditions. However, these mutants were more sensitive toward mupirocin and were impaired in survival when essential amino acids were depleted from the medium. RSH is the major enzyme responsible for (p)ppGpp synthesis in response to amino acid deprivation (lack of Leu/Val) or mupirocin treatment. Transcriptional analysis showed that the RSH-dependent stringent control in S. aureus is characterized by repression of genes whose products are predicted to be involved in the translation machinery and by upregulation of genes coding for enzymes involved in amino acid metabolism and transport which are controlled by the repressor CodY. Amino acid starvation also provoked stabilization of the RNAs coding for major virulence regulators, such as SaeRS and SarA, independently of RSH. In an animal model, the rsh syn mutant was shown to be less virulent than the wild type. Virulence could be restored by the introduction of a codY mutation into the rsh syn mutant. These results indicate that stringent conditions are present during infection and that RSH-dependent derepression of CodY-regulated genes is essential for virulence in S. aureus.

scholarly journals PheP, a Putative Amino Acid Permease of Staphylococcus aureus, Contributes to Survival In Vivo and during Starvation

2004 ◽  
Vol 72 (5) ◽  
pp. 3073-3076 ◽  
Author(s):  
Malcolm J. Horsburgh ◽  
Michael D. Wiltshire ◽  
Howard Crossley ◽  
Eileen Ingham ◽  
Simon J. Foster
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Genetic Complementation ◽  
Anaerobic Incubation ◽  
Amino Acid Permease ◽  
Poor Growth ◽  
Starvation Survival ◽  
Pig Serum

ABSTRACT PheP, a putative amino acid permease in Staphylococcus aureus, contributes to starvation survival under glucose-limiting conditions and virulence. A pheP mutation led to poor growth after microaerobic or anaerobic incubation on pig serum agar, which was recovered by phenylalanine addition. Genetic complementation of pheP restored growth and starvation survival.

scholarly journals The Role of Amino Acid Substitution in HepT Toward Menaquinone Isoprenoid Chain Length Definition and Lysocin E Sensitivity in Staphylococcus aureus

2020 ◽  
Vol 11 ◽  
Author(s):  
Suresh Panthee ◽  
Atmika Paudel ◽  
Hiroshi Hamamoto ◽  
Anne-Catrin Uhlemann ◽  
Kazuhisa Sekimizu
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Chain Length ◽  
Amino Acid Substitution
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