Characterization of novel LPXTG-containing proteins of Staphylococcus aureus identified from genome sequences

Roche, Fiona; Massey, Ruth C.; Peacock, Sharon J.; Day, Nicholas P. J.; Visai, Livia; Speziale, Pietro; Lam, Adam; Pallen, Mark J.; Foster, Timothy J.

doi:10.1099/mic.0.25996-0

Cited by 187 publications

(177 citation statements)

References 42 publications

(49 reference statements)

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“…The sequences of SasG (and Aap) repeats are very similar at the DNA level (16) and vary in number dependent on the bacterial strain (16,17). However, although sequence identity at the DNA level might be advantageous in facilitating recombination events, the resulting protein sequence identity is a potential problem from a protein folding perspective.…”

Section: Discussionmentioning

confidence: 94%

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Staphylococcal biofilm-forming protein has a contiguous rod-like structure

Gruszka

Wojdyla

Bingham

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Staphylococcus aureus and Staphylococcus epidermidis form communities (called biofilms) on inserted medical devices, leading to infections that affect many millions of patients worldwide and cause substantial morbidity and mortality. As biofilms are resistant to antibiotics, device removal is often required to resolve the infection. Thus, there is a need for new therapeutic strategies and molecular data that might assist their development. Surface proteins S. aureus surface protein G (SasG) and accumulation-associated protein ( S. epidermidis ) promote biofilm formation through their “B” regions. B regions contain tandemly arrayed G5 domains interspersed with approximately 50 residue sequences (herein called E) and have been proposed to mediate intercellular accumulation through Zn 2+ -mediated homodimerization. Although E regions are predicted to be unstructured, SasG and accumulation-associated protein form extended fibrils on the bacterial surface. Here we report structures of E–G5 and G5–E–G5 from SasG and biophysical characteristics of single and multidomain fragments. E sequences fold cooperatively and form interlocking interfaces with G5 domains in a head-to-tail fashion, resulting in a contiguous, elongated, monomeric structure. E and G5 domains lack a compact hydrophobic core, and yet G5 domain and multidomain constructs have thermodynamic stabilities only slightly lower than globular proteins of similar size. Zn 2+ does not cause SasG domains to form dimers. The work reveals a paradigm for formation of fibrils on the 100-nm scale and suggests that biofilm accumulation occurs through a mechanism distinct from the “zinc zipper.” Finally, formation of two domains by each repeat (as in SasG) might reduce misfolding in proteins when the tandem arrangement of highly similar sequences is advantageous.

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Section: Discussionmentioning

confidence: 94%

“…SasG and Aap have sequence identities of approximately 34% and approximately 50% for G5 and E repeats, respectively, and contain a variable number of repeats [three to 10 in SasG (16) and four to 17 in Aap (17)], dependent on the strain. High DNA sequence identity results in pairwise protein sequence similarity between B repeats within SasG (Fig.…”

mentioning

confidence: 99%

Staphylococcal biofilm-forming protein has a contiguous rod-like structure

Gruszka

Wojdyla

Bingham

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…We found that (i) the presence of a catheter is required for persistent MRSA UTI; (ii) MRSA binds to regions of the catheter that are coated with Fg in both mice and humans; (iii) MRSA actively promotes catheter-induced inflammation to stimulate the accumulation of Fg; and (iv) MRSA can persist during long-term catheterization despite the presence of a robust inflammatory response. Finally, analysis of the two bestcharacterized Fg-binding MSCRAMMs, Clumping Factor A (ClfA) and Clumping Factor B (ClfB) (32,(39)(40)(41), revealed ClfB was important for CAUTI, supporting a mechanism whereby MRSA exploits the presence of Fg to cause persistent CAUTI.…”

Section: Significancementioning

confidence: 99%

“…and (iii) pathogenesis in diverse animal models (28,29,(31)(32)(33)(39)(40)(41). Mutation of either gene results in decreased Fg binding and reduced virulence, albeit ClfA − strains typically display more striking phenotypes both in vitro and in vivo.…”

Section: Clfa and Clfb Mutant Strains Display Expected Fibrinogen-binmentioning

confidence: 99%

“…EbpA binding to Fg facilitates E. faecalis biofilm formation on the catheter, which initiates the CAUTI pathogenic cascade. While MRSA does not encode pili, the pathogen uses a large family of structurally homologous, cell wall-linked adhesin proteins, termed "microbial surface components recognizing adhesive matrix molecules" (MSCRAMMs), to interact with Fg and other host proteins to promote pathogenesis (32,(39)(40)(41).…”

Section: Significancementioning

confidence: 99%

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Catheterization alters bladder ecology to potentiate Staphylococcus aureus infection of the urinary tract

Walker

Flores-Mireles

Pinkner

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Methicillin-resistant Staphylococcus aureus (MRSA) is an emerging cause of catheter-associated urinary tract infection (CAUTI), which frequently progresses to more serious invasive infections. We adapted a mouse model of CAUTI to investigate how catheterization increases an individual's susceptibility to MRSA UTI. This analysis revealed that catheterization was required for MRSA to achieve high-level, persistent infection in the bladder. As shown previously, catheter placement induced an inflammatory response resulting in the release of the host protein fibrinogen (Fg), which coated the bladder and implant. Following infection, we showed that MRSA attached to the urothelium and implant in patterns that colocalized with deposited Fg. Furthermore, MRSA exacerbated the host inflammatory response to stimulate the additional release and accumulation of Fg in the urinary tract, which facilitated MRSA colonization. Consistent with this model, analysis of catheters from patients with S. aureus-positive cultures revealed colocalization of Fg, which was deposited on the catheter, with S. aureus. Clumping Factors A and B (ClfA and ClfB) have been shown to contribute to MRSA-Fg interactions in other models of disease. We found that mutants in clfA had significantly greater Fg-binding defects than mutants in clfB in several in vitro assays. Paradoxically, only the ClfB − strain was significantly attenuated in the CAUTI model. Together, these data suggest that catheterization alters the urinary tract environment to promote MRSA CAUTI pathogenesis by inducing the release of Fg, which the pathogen enhances to persist in the urinary tract despite the host's robust immune response.host-pathogen interactions | MRSA CAUTI | ClfB-fibrinogen interactions

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