An Alternative Terminal Step of the General Secretory Pathway in Staphylococcus aureus

Craney, Arryn; Dix, Melissa M.; Adhikary, Ramkrishna; Cravatt, Benjamin F.; Romesberg, Floyd E.

doi:10.1128/mbio.01178-15

Cited by 13 publications

(22 citation statements)

References 25 publications

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“…It is anchored by its C‐terminal region and has an LPXTG sortase motif. Signal peptidase cleavage adjacent to a typical AXA (ANA↓AQH) site removes a 36‐residue signal peptide as previously reported . In the current work we observed the succeeding peptide AQHDEAQQNAFYQVLNMPNLNADQR 28 times and it was amidinated 89% of the time.…”

Section: Resultssupporting

confidence: 81%

“…Distinct N‐terminal leader (signal) peptide sequences serve as determinants for one protein secretory pathway . However, there is growing evidence of other secretory pathways that involve novel processing sites typically at or near protein N‐termini . Covalent chemical labeling experiments exploit the accessibility of these sites to identify their locations.…”

Section: Introductionmentioning

confidence: 99%

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Identification of N‐terminal protein processing sites by chemical labeling mass spectrometry

Misal

Tang

et al. 2019

Rapid Comm Mass Spectrometry

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Rationale:Proteins undergo post-translational modifications and proteolytic processing that can affect their biological function. Processing often involves the loss of single residues. Cleavage of signal peptides from the N-terminus is commonly associated with translocation. Recent reports have suggested that other processing sites also exist. Methods:The secreted proteins from S. aureus N315 were precipitated with trichloroacetic acid (TCA) and amidinated with S-methyl thioacetimidate (SMTA).Amidinated proteins were digested with trypsin and analyzed with a high-resolution orbitrap mass spectrometer. Results:Sixteen examples of Staphylococcus aureus secretory proteins that lose an N-terminal signal peptide during their export were identified using this amidination approach. The N-termini of proteins with and without methionine were identified.Unanticipated protein cleavages due to sortase and an unknown protease were also uncovered. Conclusions:A simple N-terminal amidination based mass spectrometry approach is described that facilitates identification of the N-terminus of a mature protein and the discovery of unexpected processing sites.

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Section: Resultssupporting

confidence: 81%

Section: Introductionmentioning

confidence: 99%

Identification of N‐terminal protein processing sites by chemical labeling mass spectrometry

Misal

Tang

et al. 2019

Rapid Comm Mass Spectrometry

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“…The Cro/CI protein shares sequence similarity with lambda phage Cro, and has transcriptional repressor activity in S. aureus 3. Loss-of-function mutations in the cro/cI locus are associated with over-expression of the 3-gene locus immediately downstream of cro/cI which encodes a putative ABC transporter 35 (see Figure 1A). Over-expression of the ABC transporter is sufficient to confer resistance to SpsB inhibitors, whereas over-expression of the transporter with an inactivated ATPase domain is not 3.…”

mentioning

confidence: 99%

“…This suggests that this resistance mechanism is ATP-dependent and possibly mediated by active transport 3. Over-expression of the ABC transporter overcomes the S. aureus lethality caused by either pharmacologic inhibition or genetic ablation of SpsB, and is associated with secretion of alternatively cleaved proteins 35 (see Figure 2 for a model). Thus, over-expression of the putative ABC transporter is both necessary and sufficient to compensate for the essentiality of S. aureus SpsB in vitro .…”

mentioning

confidence: 99%

“…Proteomic studies of the secretome of a wild-type S. aureus strain and an S. aureus strain that lacks SpsB while over-expressing the ABC transporter show that only a minor subset of proteins that are normally SpsB-cleavable is alternatively secreted 35. These proteins, which are secreted independently of SpsB, are cleaved at a site N-terminal to the canonical SpsB-cleavage site 3.…”

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confidence: 99%

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Staphylococcus aureus type I signal peptidase: essential or not essential, that’s the question

Hazenbos¹,

Skippington²,

Tan³

2017

Microb Cell

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Secretion of proteins into the extracellular environment is crucial for the normal physiology and virulence of pathogenic bacteria. Type I signal peptidase (SPase I) mediates the final step of bacterial secretion, by cleaving proteins at their signal peptide once they are translocated by the Sec or twin-arginine (Tat) translocon. SPase I has long been thought to be essential for viability in multiple bacterial pathogens. Challenging this view, we and others have recently created Staphylococcus aureus bacteria lacking the SPase I SpsB that are viable and able to grow in vitro when over-expressing a native gene cassette encoding for a putative ABC transporter. This transporter apparently compensates for SpsB's essential function by mediating alternative cleavage of a subset of proteins at a site distinct from the SpsB-cleavage site, leading to SpsB-independent secretion. This alternative secretion system also drives the main mechanism of resistance to an arylomycin-derived SpsB inhibitor, by means of mutations in a putative transcriptional repressor (cro/cI) causing over-expression of the ABC transporter. These findings raise multiple interesting biological questions. Unraveling the mechanism of SpsB-independent secretion may provide an interesting twist to the paradigm of bacterial secretion.

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Single Atom Engineered Antibiotics Overcome Bacterial Resistance

Panáček,

Belza,

Hochvaldová

et al. 2024

Advanced Materials

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The outbreak of antibiotic‐resistant bacteria, or “superbugs”, poses a global public health hazard due to their resilience against the most effective last‐line antibiotics. Identifying potent antibacterial agents capable of evading bacterial resistance mechanisms represents the ultimate defense strategy. This study shows that –the otherwise essential micronutrient– manganese turns into a broad‐spectrum potent antibiotic when coordinated with a carboxylated nitrogen‐doped graphene. This antibiotic material (termed NGA‐Mn) not only inhibits the growth of a wide spectrum of multidrug‐resistant bacteria but also heals wounds infected by bacteria in vivo and, most importantly, effectively evades bacterial resistance development. NGA‐Mn exhibits up to 25‐fold higher cytocompatibility to human cells than its minimum bacterial inhibitory concentration, demonstrating its potential as a next‐generation antibacterial agent. Experimental findings suggest that NGA‐Mn acts on the outer side of the bacterial cell membrane via a multimolecular collective binding, blocking vital functions in both Gram‐positive and Gram‐negative bacteria. The results underscore the potential of single‐atom engineering toward potent antibiotics, offering simultaneously a long‐sought solution for evading drug resistance development while being cytocompatible to human cells.

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An Alternative Terminal Step of the General Secretory Pathway in Staphylococcus aureus

Cited by 13 publications

References 25 publications

Identification of N‐terminal protein processing sites by chemical labeling mass spectrometry

Identification of N‐terminal protein processing sites by chemical labeling mass spectrometry

Staphylococcus aureus type I signal peptidase: essential or not essential, that’s the question

Single Atom Engineered Antibiotics Overcome Bacterial Resistance

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