Comprehensive Characterization of Methicillin-resistant Staphylococcus aureus subsp. aureus COL Secretome by Two-dimensional Liquid Chromatography and Mass Spectrometry

Ravipaty, Shobha; Reilly, James P.

doi:10.1074/mcp.m900494-mcp200

Cited by 59 publications

(69 citation statements)

References 88 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In B. subtilis, the capacity of the secretion apparatus is directly dependent on PrsA (40), with ␣-amylase, protective agent, and PBPs specifically identified as PrsA substrates (41). Although numerous studies have attempted to define the S. aureus secretome, the subset of Sec exported proteins is a missing piece (42)(43)(44). PBP profiles are not affected by deletion of the secDF accessory factors of the Sec machinery (45); nevertheless, PBP3 is 1 of the 46 proteins affected by the inhibition of type 1 signal peptidase (21), and the SOSUI algorithm predicts N-terminal signal sequences for PBP4 and PBP2A (46).…”

Section: Discussionmentioning

confidence: 99%

The Staphylococcus aureus Chaperone PrsA Is a New Auxiliary Factor of Oxacillin Resistance Affecting Penicillin-Binding Protein 2A

Jousselin

Manzano

Biette

et al. 2016

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Expression of the methicillin-resistant S. aureus (MRSA) phenotype results from the expression of the extra penicillin-binding protein 2A (PBP2A), which is encoded by mecA and acquired horizontally on part of the SCCmec cassette. PBP2A can catalyze DD-transpeptidation of peptidoglycan (PG) because of its low affinity for ␤-lactam antibiotics and can functionally cooperate with the PBP2 transglycosylase in the biosynthesis of PG. Here, we focus upon the role of the membrane-bound PrsA foldase protein as a regulator of ␤-lactam resistance expression. Deletion of prsA altered oxacillin resistance in three different SCCmec backgrounds and, more importantly, caused a decrease in PBP2A membrane amounts without affecting mecA mRNA levels. The N-and C-terminal domains of PrsA were found to be critical features for PBP2A protein membrane levels and oxacillin resistance. We propose that PrsA has a role in posttranscriptional maturation of PBP2A, possibly in the export and/or folding of newly synthesized PBP2A. This additional level of control in the expression of the mecA-dependent MRSA phenotype constitutes an opportunity to expand the strategies to design anti-infective agents.

show abstract

Section: Discussionmentioning

confidence: 99%

The Staphylococcus aureus Chaperone PrsA Is a New Auxiliary Factor of Oxacillin Resistance Affecting Penicillin-Binding Protein 2A

Jousselin

Manzano

Biette

et al. 2016

Antimicrob Agents Chemother

View full text Add to dashboard Cite

show abstract

“…Furthermore, the fates of the respective Nterminal signal peptide fragments remain unknown. Notably, the studies by Ravipaty and Reilly also showed that, in certain cases, signal peptide hydrolysis within or at the membrane of S. aureus may not be fully effective, as five full-length signal peptides were also identified in the growth medium (122). These excreted signal peptides belonged to the secreted proteins Sle1, SACOL0723, SceD, IsaA, and SACOL2295.…”

Section: Signal Peptide Hydrolases Degrade Signal Peptides Within or mentioning

confidence: 99%

“…Accordingly, only a limited number of endogenous substrates have been identified for particular SPPs. Recently, in-depth proteomic studies by Ravipaty and Reilly identified signal peptide fragments of 18 secreted S. aureus proteins (122). Specifically, these were C-terminal signal peptide portions that were generated through consecutive cleavage by SPI at the signal peptidase cleavage site and by an unidentified protease within the H region.…”

Section: Signal Peptide Hydrolases Degrade Signal Peptides Within or mentioning

confidence: 99%

Membrane Proteases in the Bacterial Protein Secretion and Quality Control Pathway

Dalbey

Wang

Dijl

2012

Microbiol Mol Biol Rev

128

129

View full text Add to dashboard Cite

SUMMARY Proteolytic cleavage of proteins that are permanently or transiently associated with the cytoplasmic membrane is crucially important for a wide range of essential processes in bacteria. This applies in particular to the secretion of proteins and to membrane protein quality control. Major progress has been made in elucidating the structure-function relationships of many of the responsible membrane proteases, including signal peptidases, signal peptide hydrolases, FtsH, the rhomboid protease GlpG, and the site 1 protease DegS. These enzymes employ very different mechanisms to cleave substrates at the cytoplasmic and extracytoplasmic membrane surfaces or within the plane of the membrane. This review highlights the different ways that bacterial membrane proteases degrade their substrates, with special emphasis on catalytic mechanisms and substrate delivery to the respective active sites.

show abstract

“…Boiling staphylococci in suspension with ionic detergent does not lead to cell lysis or leakage of cytoplasmic protein, suggesting that staphylococcal murein sacculi are impenetrable for proteins (32). Nevertheless, during exponential growth, S. aureus secretes at least 59 proteins processed from signal peptide-bearing precursors into the culture medium in addition to excreting 53 polypeptides that apparently do not travel via the Sec pathway (33)(34)(35). S. aureus genes for cell wall synthesis and peptidoglycan processing were heretofore not reported to contribute to protein secretion.…”

mentioning

confidence: 99%

SagB Glucosaminidase Is a Determinant of Staphylococcus aureus Glycan Chain Length, Antibiotic Susceptibility, and Protein Secretion

et al. 2016

View full text Add to dashboard Cite

The envelope of Staphylococcus aureus is comprised of peptidoglycan and its attached secondary polymers, teichoic acid, capsular polysaccharide, and protein. Peptidoglycan synthesis involves polymerization of lipid II precursors into glycan strands that are cross-linked at wall peptides. It is not clear whether peptidoglycan structure is principally determined during polymerization or whether processive enzymes affect cell wall structure and function, for example, by generating conduits for protein secretion. We show here that S. aureus lacking SagB, a membrane-associated N-acetylglucosaminidase, displays growth and cell-morphological defects caused by the exaggerated length of peptidoglycan strands. SagB cleaves polymerized glycan strands to their physiological length and modulates antibiotic resistance in methicillin-resistant S. aureus (MRSA). Deletion of sagB perturbs protein trafficking into and across the envelope, conferring defects in cell wall anchoring and secretion, as well as aberrant excretion of cytoplasmic proteins. IMPORTANCEStaphylococcus aureus is thought to secrete proteins across the plasma membrane via the Sec pathway; however, protein transport across the cell wall envelope has heretofore not been studied. We report that S. aureus sagB mutants generate elongated peptidoglycan strands and display defects in protein secretion as well as aberrant excretion of cytoplasmic proteins. These results suggest that the thick peptidoglycan layer of staphylococci presents a barrier for protein secretion and that SagB appears to extend the Sec pathway across the cell wall envelope. Staphylococcus aureus, a Gram-positive bacterial pathogen, replicates via septal assembly of membranes and peptidoglycan into the cross wall compartment (1, 2). The peptidoglycan of the cross wall is split by murein hydrolases, separating daughter cells that assume a spherical shape (3). Earlier work identified three murein hydrolases with cross-wall-splitting activities: Atl (autolysin), Sle1, and LytN (3-5). Atl and Sle1 are secreted into the extracellular milieu and subsequently cleave septal peptidoglycan at the cross wall but not elsewhere as access is restricted by teichoic acid modification of peptidoglycan (6-8). LytN, on the other hand, is secreted into the cross wall compartment (5). S. aureus Atl is synthesized as a preproenzyme with an N-terminal signal peptide and prodomain (9, 10). Secreted pro-Atl is processed to generate Atl N-acetylmuramoyl-L-Ala-amidase (Atl AM ) and Atl Nacetylglucosaminidase (Atl GL ), and each binds via GW domains to lipoteichoic acids (10-12). Earlier work demonstrated that Atl functions as an endo-␤-N-acetylglucosaminidase (12, 13). Although initially designated autolysin (Atl), the S. aureus atl mutant does not display an autolysis phenotype yet forms large clusters of incompletely separated bacteria and is defective for penicillin-induced killing (14). The LysM domains of Sle1 promote its binding to cross wall peptidoglycan, and sle1 mutants also form clusters of incompletely s...

show abstract

Comprehensive Characterization of Methicillin-resistant Staphylococcus aureus subsp. aureus COL Secretome by Two-dimensional Liquid Chromatography and Mass Spectrometry

Cited by 59 publications

References 88 publications

The Staphylococcus aureus Chaperone PrsA Is a New Auxiliary Factor of Oxacillin Resistance Affecting Penicillin-Binding Protein 2A

The Staphylococcus aureus Chaperone PrsA Is a New Auxiliary Factor of Oxacillin Resistance Affecting Penicillin-Binding Protein 2A

Membrane Proteases in the Bacterial Protein Secretion and Quality Control Pathway

SagB Glucosaminidase Is a Determinant of Staphylococcus aureus Glycan Chain Length, Antibiotic Susceptibility, and Protein Secretion

Contact Info

Product

Resources

About