A Study on the Biosynthesis of Protein A in <i>Staphylococcus aureus</i>

Movitz, Jan

doi:10.1111/j.1432-1033.1974.tb03750.x

Cited by 26 publications

(12 citation statements)

References 18 publications

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“…5B, treatment of staphylococci with penicillin, vancomycin, bacitracin, or moenomycin led to a significant reduction in the amount of protein A incorporated into the cell wall. Vancomycin and moenomycin caused the largest decrease in surface protein anchoring and display, consistent with previous reports that these antibiotics prevent the incorporation of polypeptides into the cell wall envelope (27,41).…”

Section: Measuring the Incorporation Of Radiolabel By S Aureus Cultusupporting

confidence: 91%

Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus

Perry

Ton‐That

Mazmanian

et al. 2002

Journal of Biological Chemistry

204

106

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Surface proteins of Staphylococcus aureus are anchored to the cell wall peptidoglycan by a mechanism requiring a C-terminal sorting signal with an LPXTG motif. Surface proteins are first synthesized in the bacterial cytoplasm and then transported across the cytoplasmic membrane. Cleavage of the N-terminal signal peptide of the cytoplasmic surface protein P1 precursor generates the extracellular P2 species, which is the substrate for the cell wall anchoring reaction. Sortase, a membrane-anchored transpeptidase, cleaves P2 between the threonine (T) and the glycine (G) of the LPXTG motif and catalyzes the formation of an amide bond between the carboxyl group of threonine and the amino group of cell wall cross-bridges. We have used metabolic labeling of staphylococcal cultures with To mount a successful infection, Gram-positive pathogens display proteins on the bacterial surface that adhere to specific receptors on host tissues or provide for microbial escape from the host's immune response (1). Protein display on the bacterial surface involves the covalent linkage of polypeptides to the cell wall envelope (2). As reported for protein A of Staphylococcus aureus, surface proteins are synthesized as P1 precursor molecules in the bacterial cytoplasm, bearing an N-terminal signal peptide and a C-terminal sorting signal (3). The 35-residue sorting signal is composed of a LPXTG motif, a hydrophobic domain, and a tail of positively charged residues (4). After translocation across the cytoplasmic membrane, the N-terminal signal peptide is removed by signal peptidase, thereby generating the P2 precursor (4). The C-terminal sorting signal retains the P2 precursor species within the secretory pathway and permits substrate recognition at the LPXTG motif (4, 5). Sortase, a membrane-anchored transpeptidase, cleaves surface proteins between the threonine (T) and the glycine (G) of the LPXTG motif (6, 7). Cleaved polypeptides are initially tethered as thioester-linked intermediates to the active site sulfhydryl residue of sortase enzymes (8). Nucleophilic attack of the amino group of pentaglycine cross-bridges within the staphylococcal peptidoglycan resolves this acyl-enzyme intermediate (8), resulting in the formation of an amide bond that tethers the C terminus of surface protein to the cell wall peptidoglycan (9 -13).The peptidoglycan of S. aureus is synthesized in three cellular compartments, the cytoplasm, the membrane and the cell wall envelope ( (18,19). Lipid II is translocated across the cytoplasmic membrane and functions as a substrate for two cell wall biosynthetic reactions that require mono-or bifunctional transglycosylases and transpeptidases (20). In the transglycosylation reaction, lipid II is polymerized to generate linear peptidoglycan strands with the repeating disaccharide (MurNAc-GlcNAc) n . This reaction is fueled by the hydrolysis of lipid II and by further hydrolysis of the undecaprenolpyrophosphate product, which is translocated across the plasma membrane into the cytoplasm (21). Linear peptidoglycan strand...

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Section: Measuring the Incorporation Of Radiolabel By S Aureus Cultusupporting

confidence: 91%

Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus

Perry

Ton‐That

Mazmanian

et al. 2002

Journal of Biological Chemistry

204

106

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“…Others, e.g., enterotoxin B, metalloprotease, and nuclease (3,7,11), are produced at very low levels throughout the growth cycle but more actively after exponential growth. In contrast, a small number of cell wall-associated proteins such as coagulase and protein A are synthesized preferentially during exponential growth (12,27,28). These observations imply that there is a coordinate and temporal control over the production of these proteins.…”

mentioning

confidence: 62%

A temporal signal, independent of agr, is required for hla but not spa transcription in Staphylococcus aureus

Vandenesch¹,

Kornblum²,

Novick³

1991

J Bacteriol

167

155

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Staphylococcus aureus exoprotein expression is controlled by a global regulon known as agr. This system activates transcription of some target genes and represses transcription of others. Target genes expressed postexponentially such as a-hemolysin (hla) are activated by agr; target genes expressed during exponential phase such as protein A (spa) are repressed by agr. A unique feature of the agr system is that this transcriptional regulation is mediated by a 517-nucleotide transcript, RNAIII. While it is clear that agr differentially regulates the expression of exponential and postexponential exoproteins, the precise role of agr in the temporal control of these events has not yet been explored. In this report, we examine the effects of expressing RNAIII, the agr regulator, under the control of the inducible ,B-lactamase (bla) promoter at different times in the growth cycle. We confirm previous results showing that agr is required for postexponential-phase expression of hla and further show that a separate postexponential-phase signal independent of agr function is also needed for activation of hla transcription. We also show that in an agr mutant transcription of spa occurs throughout the growth cycle, is inhibited immediately upon induction of RNAIII, and is thus indifferent to the postexponential signal required for hia activation.

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“…vitz (38) examined the effect of vancomycin on the incorporation of protein A into cell wall polymer and observed no effect. In contrast to our measurements of the rate of cell wall sorting precursor cleavage, these observations relied on the binding of cell wall-associated protein A to immunoglobulin as a measure of the amount of surface protein anchoring.…”

Section: Discussionmentioning

confidence: 99%

Anchor Structure of Staphylococcal Surface Proteins

Ton‐That

Schneewind

1999

Journal of Biological Chemistry

140

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Surface proteins of Staphylococcus aureus are covalently linked to the bacterial cell wall by a mechanism requiring a COOH-terminal sorting signal with a conserved LPXTG motif. Cleavage between the threonine and the glycine of the LPXTG motif liberates the carboxyl of threonine to form an amide bond with the amino of the pentaglycine cross-bridge in the staphylococcal peptidoglycan. We asked whether antibiotic cell wall synthesis inhibitors interfere with the anchoring of surface proteins. Penicillin G, a transpeptidation inhibitor, had no effect on surface protein anchoring, whereas vancomycin and moenomycin, inhibitors of cell wall polymerization into peptidoglycan strands, slowed the sorting reaction. Cleavage of surface protein precursors did not require a mature assembled cell wall and was observed in staphylococcal protoplasts. A search for chemical inhibitors of the sorting reaction identified methanethiosulfonates and p-hydroxymercuribenzoic acid. Thus, sortase, the enzyme proposed to cleave surface proteins at the LPXTG motif, appears to be a sulfhydryl-containing enzyme that utilizes peptidoglycan precursors but not an assembled cell wall as a substrate for the anchoring of surface protein.

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A Study on the Biosynthesis of Protein A in Staphylococcus aureus

Cited by 26 publications

References 18 publications

Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus

Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus

A temporal signal, independent of agr, is required for hla but not spa transcription in Staphylococcus aureus

Anchor Structure of Staphylococcal Surface Proteins

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