The Intracellular Cyclophilin PpiB Contributes to the Virulence of Staphylococcus aureus Independently of Its Peptidyl-Prolyl
            <i>cis/trans</i>
            Isomerase Activity

Keogh, Rebecca A.; Zapf, Rachel L.; Wiemels, Richard E.; Wittekind, Marcus A.; O’Carroll, Ronan

doi:10.1128/iai.00379-18

Cited by 17 publications

(29 citation statements)

References 46 publications

(52 reference statements)

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“…Murine model of skin abscess formation. The murine model of skin abscess formation was performed as described previously (66). Briefly, wild-type and mutant strains were grown for 2.5 h until reaching an OD 600 of approximately 0.75.…”

Section: Methodsmentioning

confidence: 99%

MroQ Is a Novel Abi-Domain Protein That Influences Virulence Gene Expression in Staphylococcus aureus via Modulation of Agr Activity

et al. 2019

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Numerous factors have, to date, been identified as playing a role in the regulation of Agr activity in Staphylococcus aureus, including transcription factors, antisense RNAs, and host elements. Herein we investigated the product of SAUSA300_ 1984 (termed MroQ), a transmembrane Abi-domain/M79 protease-family protein, as a novel effector of this system. Using a USA300 mroQ mutant, we observed a drastic reduction in proteolysis, hemolysis, and pigmentation that was fully complementable. This appears to result from diminished agr activity, as transcriptional analysis revealed significant decreases in expression of both RNAII and RNAIII in the mroQ mutant. Such effects appear to be direct, rather than indirect, as known agr effectors demonstrated limited alterations in their activity upon mroQ disruption. A comparison of RNA sequencing data sets for both mroQ and agr mutants revealed a profound overlap in their regulomes, with the majority of factors affected being known virulence determinants. Importantly, the preponderance of alterations in expression were more striking in the agr mutant, indicating that MroQ is necessary, but not sufficient, for Agr function. Mechanism profiling revealed that putative residues for metalloprotease activity within MroQ are required for its Agr-controlling effect; however, this was not wielded at the level of AgrD processing. Virulence assessment demonstrated that both mroQ and agr mutants exhibited increased formation of renal abscesses but decreased skin abscess formation alongside diminished dermonecrosis. Collectively, we present the characterization of a novel agr effector in S. aureus which would appear to be a direct regulator, potentially functioning via interaction with the AgrC histidine kinase.

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Section: Methodsmentioning

confidence: 99%

MroQ Is a Novel Abi-Domain Protein That Influences Virulence Gene Expression in Staphylococcus aureus via Modulation of Agr Activity

et al. 2019

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“…These experiments were performed as described previously (68). Briefly, wild type and mutant strains were grown for 2.5 h until reaching an OD 600 of approximately 0.75.…”

Section: Methodsmentioning

confidence: 99%

MroQ is a Novel Abi-domain Protein That Influences Virulence Gene Expression in Staphylococcus aureus via Modulation of Agr Activity

Marroquin

Gimza

Tomlinson

et al. 2019

Preprint

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17Numerous factors have to date been identified as playing a role in the regulation of Agr 18 activity in S. aureus, including transcription factors, antisense RNAs, and host elements. 19Herein we investigate the product of SAUSA300_1984 (termed MroQ), a transmembrane 20Abi-domain/M79 protease-family protein, as a novel effector of this system. Using a 21 USA300 mroQ mutant we observed a drastic reduction in proteolysis, hemolysis and 22 pigmentation that was fully complementable. This appears to result from diminished agr 23 activity, as transcriptional analysis revealed significant decreases in expression of both 24 RNAII and RNAIII in the mroQ mutant. Such effects appear to be direct, rather than 25 indirect, as known agr effectors demonstrated limited alterations in their activity upon 26 mroQ disruption. A comparison of RNA-sequencing datasets for both mroQ and agr 27 mutants reveal a profound overlap in their regulomes, with the majority of factors affected 28 being known virulence determinants. Importantly, the preponderance of alterations in 29 expression were more striking in the agr mutant, indicating that MroQ is necessary, but 30 not sufficient, for Agr function. Mechanism profiling revealed that putative residues for 31 metalloprotease activity within MroQ are required for its Agr controlling effect, however 32 this is not wielded at the level of AgrD processing. Virulence assessment demonstrated 33 that mroQ and agr mutants both exhibited increased formation of renal abscesses, but 34 decreased skin abscess formation, alongside diminished dermonecrosis. Collectively, we 35 present the characterization of a novel agr effector in S. aureus, which would appear to 36 be a direct regulator, potentially functioning via interaction with the AgrC histidine kinase. 37Pathogenicity in S. aureus is both diverse and tightly controlled, being highly adaptive to 39 the surrounding environment and available resources (1). In order to survive and persist 40 within the host, this pathogen produces a wide array of virulence factors throughout the 41 different stages of infection (2). For example, during earlier phases of growth, adhesins 42 and other binding proteins are actively synthesized (e.g. Spa, FnbAB), which play a role 43 in colonization (2, 3). In contrast, in later phases, these surface proteins are repressed, 44 whilst secreted toxins, proteases, and exoenzymes are produced to facilitate invasion and 45 dissemination (2, 4). This coordination is facilitated by an array of global regulators of 46 gene expression (2, 5-10). 47 48The agr quorum sensing system is one of the most important regulators of virulence in S. 49 aureus (11). It is a quorum sensing system comprised of two transcriptional units, RNAII 50 and RNAIII, each controlled by its own promoter, P2 and P3, respectively. RNAII encodes 51 the agr operon, which is comprised of four genes, agrBDCA (12). The quorum sensing 52 system functions in three parts: i) a quorum sensing module comprised of AgrB and AgrD; 53ii) a two-component system (TCS) Agr...

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“…aureus -encoded virulence factor seems to be folded by SaCyp [46]. Interestingly, a point mutation in SaCyp that abolished its PPIase activity partly refolded a denatured nuclease [47]. An S .…”

Section: Introductionmentioning

confidence: 99%

“…An S . aureus mutant carrying the above mutation also exhibited pathogenicity in mouse and possessed hemolytic activity [47]. Jointly, SaCyp could be exploited in developing and screening the novel antistaphylococcal agents capable of preventing the infections caused by the multidrug-resistant S .…”

Section: Introductionmentioning

confidence: 99%

A staphylococcal cyclophilin carries a single domain and unfolds via the formation of an intermediate that preserves cyclosporin A binding activity

Seal

Polley

2019

PLoS ONE

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Cyclophilin (Cyp), a peptidyl-prolyl cis - trans isomerase (PPIase), acts as a virulence factor in many bacteria including Staphylococcus aureus . The enzymatic activity of Cyp is inhibited by cyclosporin A (CsA), an immunosuppressive drug. To precisely determine the unfolding mechanism and the domain structure of Cyp, we have investigated a chimeric S . aureus Cyp (rCyp) using various probes. Our limited proteolysis and the consequent analysis of the proteolytic fragments indicate that rCyp is composed of one domain with a short flexible tail at the C-terminal end. We also show that the urea-induced unfolding of both rCyp and rCyp-CsA is completely reversible and proceeds via the synthesis of at least one stable intermediate. Both the secondary structure and the tertiary structure of each intermediate appears very similar to those of the corresponding native protein. Conversely, the hydrophobic surface areas of the intermediates are comparatively less. Further analyses reveal no loss of CsA binding activity in rCyp intermediate. The thermodynamic stability of rCyp was also significantly increased in the presence of CsA, recommending that this protein could be employed to screen new CsA derivatives in the future.

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The Intracellular Cyclophilin PpiB Contributes to the Virulence of Staphylococcus aureus Independently of Its Peptidyl-Prolyl cis/trans Isomerase Activity

Cited by 17 publications

References 46 publications

MroQ Is a Novel Abi-Domain Protein That Influences Virulence Gene Expression in Staphylococcus aureus via Modulation of Agr Activity

MroQ Is a Novel Abi-Domain Protein That Influences Virulence Gene Expression in Staphylococcus aureus via Modulation of Agr Activity

MroQ is a Novel Abi-domain Protein That Influences Virulence Gene Expression in Staphylococcus aureus via Modulation of Agr Activity

A staphylococcal cyclophilin carries a single domain and unfolds via the formation of an intermediate that preserves cyclosporin A binding activity

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