The SrtA Sortase of
            <i>Streptococcus agalactiae</i>
            Is Required for Cell Wall Anchoring of Proteins Containing the LPXTG Motif, for Adhesion to Epithelial Cells, and for Colonization of the Mouse Intestine

Lalioui, Lila; Pellegrini, Elisabeth; Dramsi, Shaynoor; Baptista, Manuela; Bourgeois, N.; Doucet‐Populaire, Florence; Rusniok, Christophe; Zouine, Mohamed; Glaser, Philippe; Kunst, Frank; Poyart, Claire; Trieu-Cuot, Patrick

doi:10.1128/iai.73.6.3342-3350.2005

Cited by 108 publications

(107 citation statements)

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“…Earlier findings have shown that the fibronectin binding ability of srtA − mutants of Staphylococcus aureus (Mazmanian et al 2000) and S. agalactiae (Lalioui et al 2005) was severely hampered compared to wild type bacteria. We screened 6 vinyl sulfone drugs (by a fibronectin-binding ELISA) for their ability to inhibit the binding of Renibacterium salmoninarum to Chinook salmon fibronectin.…”

Section: Discussionmentioning

confidence: 99%

Sortase inhibitor phenyl vinyl sulfone inhibits Renibacterium salmoninarum adherence and invasion of host cells

Sudheesh¹,

Crane²,

Cain³

et al. 2007

Dis. Aquat. Org.

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Renibacterium salmoninarum, the causative agent of bacterial kidney disease in salmonid fishes, is a Gram-positive diplococcobacillus belonging to the family Micrococcaceae. Analysis of the genome sequence of the bacterium demonstrated the presence of a sortase homolog (srtD), a gene specifying an enzyme found in Gram-positive bacteria and required for covalent anchoring of cell surface proteins. Interference of sortase activity is being examined as a target for therapeutic prevention of infection by several pathogenic Gram-positive bacterial species. In silico analysis identified 8 open reading frames containing sortase recognition motifs, suggesting these proteins are translocated to the bacterial cell wall. The sortase and potential sortase substrate genes are transcribed in R. salmoninarum, suggesting they encode functional proteins. Treatment of R. salmoninarum with phenyl vinyl sulfone (PVS) significantly reduced bacterial adherence to Chinook salmon fibronectin. In addition, the ability of the PVS-treated bacteria to adhere to Chinook salmon embryo cells in vitro was dramatically reduced compared to that of untreated bacteria. More importantly, PVS-treated bacteria were unable to invade and replicate within CHSE-214 cells (demonstrated by an intracellular growth assay and by light microscopy). When treated with PVS, R. salmoninarum was not cytopathic to CHSE-214 cells, whereas untreated bacteria produced cytopathology within a few days. These findings clearly show that PVS, a small molecule drug and a known sortase inhibitor, can interfere with the ability of R. salmoninarum to adhere and colonize fish cells, with a corresponding decrease in virulence.KEY WORDS: Renibacterium salmoninarum · Bacterial kidney disease · Anti-virulence chemotherapy · Adherence · Invasion · Host cell sortase inhibitor · Phenyl vinyl sulfone Resale or republication not permitted without written consent of the publisherDis Aquat Org 78: [115][116][117][118][119][120][121][122][123][124][125][126][127] 2007 1994, Kehoe 1994). MSCRAMs have specific C-terminal anchoring signals or cell wall sorting (CWS)-motifs that are recognized by a membrane transpeptidase called sortase. Sortase catalyzes covalent anchoring of these proteins to the peptidoglycan surface during cell wall synthesis (reviewed in Navarre & Schneewind 1999, Ton-That et al. 2004, Marraffini et al. 2006). Elimination of sortase activity and the consequent disruption of surface protein anchoring correlates with a dramatic decrease in pathogenicity of Gram-positive bacteria in animal infections (Mazmanian et al. 2000, Bolken et al. 2001, Bierne et al. 2002, Garandeau et al. 2002, Jonsson et al. 2002, Weiss et al. 2004. Sortase is also involved in the assembly of cell surface pili on Gram-positive bacteria, which often aid attachment to host cells (Ton-That & Schneewind 2003). Collectively, these and other findings suggest that sortases are promising targets for new antibacterial drugs (Schneewind et al. 1993, Frankel et al. 2004, Ton-That et al. 2004, Weis...

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Section: Discussionmentioning

confidence: 99%

Sortase inhibitor phenyl vinyl sulfone inhibits Renibacterium salmoninarum adherence and invasion of host cells

Sudheesh¹,

Crane²,

Cain³

et al. 2007

Dis. Aquat. Org.

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“…Pili are formed from polymerized proteins, which have an Nterminal conserved signal peptide and a C-terminal LPXTG motif; the LPXTG domain enabling covalent attachment of the pilus proteins to the cell-wall peptidoglycan (Lalioui et al, 2005;Nobbs et al, 2008;Ton-That & Schneewind, 2004). GBS pili have been described as contributing to GBS adhesiveness and supporting the transcytoses of the bacteria through differentiated human epithelial cells (Pezzicoli et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Distribution of pilus islands of group B streptococcus associated with maternal colonization and invasive disease in South Africa

Madzivhandila

Adrian

Cutland

et al. 2013

Journal of Medical Microbiology

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Group B streptococcus (GBS) is a leading cause of neonatal sepsis. Sortase-dependent piluslike structures have been identified on the surface of GBS, and have been found to be important in the adhesion and attachment of GBS to host cells. Three pilus island alleles, PI-1, PI-2a and PI2b, have been described, and their proteins are being explored as vaccine candidates. The pilus islands from 541 colonization isolates and 284 invasive isolates were characterized by PCR. All isolates carried at least one pilus island, and they were identified alone or in combinations at the following overall frequencies: PI-2a, 29.8 %; PI-2b, 0.2 %; PI-1+PI-2a, 24.8 %; and PI-1+PI-2b, 45.1 %. A combination of PI-1+PI-2a (28.7 vs 17.6 %) was more common among colonizing compared with invasive isolates. Conversely, a combination of PI-1+PI-2b (37.2 vs 60.2 %) was more frequently associated with invasive disease compared to colonization. There was a strong association between pilus islands when adjusted for serotype distribution, PI-2a was identified in 92.6 % of colonizing and 90.0 % of invasive serotype Ia isolates, whereas serotype III was associated with co-expression of a PI-1 and PI-2b among 84.6 % of colonizing and 96.5 % of invasive isolates. Based on this homogeneity of pilus island distribution, a pilus-based vaccine developed for Europe and the USA will have similar coverage in South Africa.

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“…In Streptococcus gordonii, srtA inactivation leads to decreased surface protein display, reduced fibronectin binding in vitro, and a reduced ability to colonize the oral mucosa of mice (10). Likewise, srtA knockouts from Streptococcus family members S. mutans (11), S. pneumonia (12), S. pyogenes (13), S. suis (14), S. sanguinis (15), and S. agalactiae (16) have been shown to have attenuated pathogenicity.…”

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confidence: 99%

Engineering the Substrate Specificity of Staphylococcus aureus Sortase A

Bentley

Gaweska

Kielec

et al. 2007

Journal of Biological Chemistry

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The Staphylococcus aureus transpeptidase Sortase A (SrtA) anchors virulence and colonization-associated surface proteins to the cell wall. SrtA selectively recognizes a C-terminal LPXTG motif, whereas the related transpeptidase Sortase B (SrtB) recognizes a C-terminal NPQTN motif. In both enzymes, cleavage occurs after the conserved threonine, followed by amide bond formation between threonine and the pentaglycine cross-bridge of cell wall peptidoglycan. Genetic and biochemical studies strongly suggest that SrtA and SrtB exhibit exquisite specificity for their recognition motifs. To better understand the origins of substrate specificity within these two isoforms, we used sequence and structural analysis to predict residues and domains likely to be involved in conferring substrate specificity. Mutational analyses and domain swapping experiments were conducted to test their function in substrate recognition and specificity. Marked changes in the specificity profile of SrtA were obtained by replacing the ␤6/␤7 loop in SrtA with the corresponding domain from SrtB. The chimeric ␤6/␤7 loop swap enzyme (SrtLS) conferred the ability to acylate NPQTNcontaining substrates, with a k cat /K m app of 0.0062 ؎ 0.003 M ؊1 s ؊1 . This enzyme was unable to perform the transpeptidation stage of the reaction, suggesting that additional domains are required for transpeptidation to occur. The overall catalytic specificity profile (k cat /K m app (NPQTN)/k cat /K m app (LPETG)) of SrtLS was altered 700,000-fold from SrtA. These results indicate that the ␤6/␤7 loop is an important site for substrate recognition in sortases.

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The SrtA Sortase of Streptococcus agalactiae Is Required for Cell Wall Anchoring of Proteins Containing the LPXTG Motif, for Adhesion to Epithelial Cells, and for Colonization of the Mouse Intestine

Cited by 108 publications

References 59 publications

Sortase inhibitor phenyl vinyl sulfone inhibits Renibacterium salmoninarum adherence and invasion of host cells

Sortase inhibitor phenyl vinyl sulfone inhibits Renibacterium salmoninarum adherence and invasion of host cells

Distribution of pilus islands of group B streptococcus associated with maternal colonization and invasive disease in South Africa

Engineering the Substrate Specificity of Staphylococcus aureus Sortase A

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