A streptococcal effector protein that inhibits Porphyromonas gingivalis biofilm development

Christopher, Aaron B.; Arndt, Annette; Cugini, Carla; Davey, Mary E.

doi:10.1099/mic.0.042671-0

Cited by 51 publications

(48 citation statements)

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“…P. gingivalis strain variation could contribute to these differences in gene expression and/or variation in gene regulation. In addition, using chemical inhibitors, they obtained data that indicated that enzymic activity was not required for ADI signalling and our early studies showed that ADI, which appeared to be inactive (unable to convert arginine to citrulline in a standard ADI assay), could inhibit P. gingivalis biofilm formation (Christopher et al, 2010;Xie et al, 2007). However, we have since determined (data presented here) that enzymic activity of ADI is sensitive to the redox state.…”

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confidence: 95%

“…Within the oral biofilm community, the release of ammonia from this first reaction plays an important role in inhibiting tooth decay and in modulating the microbial composition of the community through neutralization of acid end products that are produced by a variety of oral bacteria via fermentation of sugars (Burne & Marquis, 2000;Casiano-Coló n & Marquis, 1988; Nascimento et al, 2009). In contrast with its canonical role in anaerobic ATP synthesis within the cytoplasm, ADI's function as an extracellular enzyme is unclear.Our studies have shown that exposure of P. gingivalis strain 381 to ADI secreted by S. intermedius results in a decrease in the expression of fimbrial subunits (encoded by fimA and mfa1), which are both key virulence determinants and essential to biofilm development (Amano et al, 2004;Christopher et al, 2010; Hamada et al, 1998;Lamont & Jenkinson, 1998;Xie et al, 2007). This interspecies inhibition is consistent with reports by Xie and colleagues that showed the effect of a cell-wall-associated Streptococcus cristatus arginine deiminase on P. gingivalis strain 33277, with the only distinction being that in strain 33277 expression of fimA, but not mfa1, is altered (Wu & Xie, 2010;Xie et al, 2007).…”

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confidence: 99%

“…Arginine is not only an immediate precursor for protein synthesis; it is also catabolized to critical cell-signalling molecules, such as nitric oxide and glutamate (Wu & Morris, 1998). Thus, we hypothesize that there is likely to be a complex interplay between host and microbiota for this substrate and that the utilization of this resource is tightly regulated.In our previous studies, we discovered an extracellular arginine deiminase (ADI) produced by the oral bacterium Streptococcus intermedius that inhibited P. gingivalis biofilm formation, without affecting its planktonic growth (Christopher et al, 2010). The canonical role of ADI is to catalyse the intracellular hydrolytic conversion of free Larginine to L-citrulline and ammonia ( Fig.…”

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confidence: 99%

“…Cultures (1 ml) were pelleted, the supernatants were carefully removed, and pellets were resuspended into 5 ml 26 CDM/1 % tryptone prior to addition to a 96-well plate (BD Falcon, no. 353936) (Christopher et al, 2010;Milner et al, 1996). Each well consisted of a total volume of 200 ml, containing 100 ml of the cell resuspension and the appropriate protein and/or buffer.…”

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confidence: 99%

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Arginine deiminase inhibits Porphyromonas gingivalis surface attachment

Cugini

Stephens²,

Nguyen³

et al. 2013

Microbiology

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The oral cavity is host to a complex microbial community whose maintenance depends on an array of cell-to-cell interactions and communication networks, with little known regarding the nature of the signals or mechanisms by which they are sensed and transmitted. Determining the signals that control attachment, biofilm development and outgrowth of oral pathogens is fundamental to understanding pathogenic biofilm development. We have previously identified a secreted arginine deiminase (ADI) produced by Streptococcus intermedius that inhibited biofilm development of the commensal pathogen Porphyromonas gingivalis through downregulation of genes encoding the major (fimA) and minor (mfa1) fimbriae, both of which are required for proper biofilm development. Here we report that this inhibitory effect is dependent on enzymic activity. We have successfully cloned, expressed and defined the conditions to ensure that ADI from S. intermedius is enzymically active. Along with the cloning of the wild-type allele, we have created a catalytic mutant (ADIC399S), in which the resulting protein is not able to catalyse the hydrolysis of L-arginine to L-citrulline. P. gingivalis is insensitive to the ADIC399S catalytic mutant, demonstrating that enzymic activity is required for the effects of ADI on biofilm formation. Biofilm formation is absent under L-arginine-deplete conditions, and can be recovered by the addition of the amino acid. Taken together, the results indicate that arginine is an important signal that directs biofilm formation by this anaerobe. Based on our findings, we postulate that ADI functions to reduce arginine levels and, by a yet to be identified mechanism, signals P. gingivalis to alter biofilm development. ADI release from the streptococcal cell and its cross-genera effects are important findings in understanding the nature of inter-bacterial signalling and biofilm-mediated diseases of the oral cavity. INTRODUCTIONPorphyromonas gingivalis is an oral pathobiont, i.e. a natural member of the human microbiota, that under certain perturbations to the host and/or microflora can cause pathology. This Gram-negative, highly proteolytic anaerobe is regarded as the primary aetiological agent of adult periodontal disease, leading to chronic inflammation and destruction of both the soft and hard tissues supporting the teeth (Choil et al., 1990;Dzink et al., 1988; Grossi et al., 1994;Lamont & Jenkinson, 2000;Moore et al., 1991). As a pathobiont, the ability to proliferate and express virulence determinants is central to its shift to pathogenicity, thus determining the mechanisms that control the emergence of its pathogenic state are of fundamental importance.P. gingivalis is a strict anaerobe that preferentially utilizes protein or peptide substrates for growth. Although studies have shown that P. gingivalis may utilize free amino acids or dipeptides, the uptake and growth rates on these substrates are limited and highly variable among strains (Seddon et al., 1988;Takahashi et al., 2000;Tang-Larsen et al., 1995). In general...

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confidence: 95%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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Arginine deiminase inhibits Porphyromonas gingivalis surface attachment

Cugini

Stephens²,

Nguyen³

et al. 2013

Microbiology

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“…To verify the fold change in expression of the identified genes, candidate genes from microarray analysis (35 genes in TSB and 48 genes under ironlimited growth conditions) were then analysed by qRT-PCR, as previously described (Christopher et al, 2010) with the following modifications. In brief, total RNA was confirmed to be free of genomic contamination by conventional PCR using the PG0521 (groES) primer and the RNA as a template.…”

Section: Methodsmentioning

confidence: 99%

The nucleoid-associated protein HUβ affects global gene expression in Porphyromonas gingivalis

et al. 2013

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HU is a non-sequence-specific DNA-binding protein and one of the most abundant nucleoidassociated proteins in the bacterial cell. Like Escherichia coli, the genome of Porphyromonas gingivalis is predicted to encode both the HUa (PG1258) and the HUb (PG0121) subunit. We have previously reported that PG0121 encodes a non-specific DNA-binding protein and that PG0121 is co-transcribed with the K-antigen capsule synthesis operon. We also reported that deletion of PG0121 resulted in downregulation of capsule operon expression and produced a P. gingivalis strain that is phenotypically deficient in surface polysaccharide production. Here, we show through complementation experiments in an E. coli MG1655 hupAB double mutant strain that PG0121 encodes a functional HU homologue. Microarray and quantitative RT-PCR analysis were used to further investigate global transcriptional regulation by HUb using comparative expression profiling of the PG0121 (HUb) mutant strain to the parent strain, W83. Our analysis determined that expression of genes encoding proteins involved in a variety of biological functions, including iron acquisition, cell division and translation, as well as a number of predicted nucleoid associated proteins were altered in the PG0121 mutant. Phenotypic and quantitative real-time-PCR (qRT-PCR) analyses determined that under iron-limiting growth conditions, cell division and viability were defective in the PG0121 mutant. Collectively, our studies show that PG0121 does indeed encode a functional HU homologue, and HUb has global regulatory functions in P. gingivalis; it affects not only production of capsular polysaccharides but also expression of genes involved in basic functions, such as cell wall synthesis, cell division and iron uptake. INTRODUCTIONPorphyromonas gingivalis is a Gram-negative obligate anaerobe belonging to the family Bacteroidaceae that persists as a natural member of the human oral microbiota. A shift in the microbial community leading to outgrowth of this anaerobe is directly linked to periodontitis, a chronic inflammatory disease that leads to destruction of the tissues supporting the gums and ultimately, exfoliation of the teeth (Choil et al., 1990;Dzink et al., 1988;Grossi et al., 1994;Lamont & Jenkinson, 2000;Moore et al., 1991). This commensal can colonize, invade and multiply within gingival epithelial cells, as well as penetrate into deeper epithelial cell layers, potentially releasing the whole organism and/or virulence factors into the bloodstream (reviewed by Yilmaz, 2008). In addition to its ability to cause disease in the oral cavity, there are data indicating a role in systemic disease, including its ability to invade vascular endothelial cells (Dorn et al., 2000(Dorn et al., , 2002Jandik et al., 2008) and to cause aggregation of platelets (Pham et al., 2002). For many pathogenic bacteria, surface polysaccharides play a key role in immune modulation et al., 1996). HU typically acts as an accessory protein in virtually all types of nucleoprotein-mediated processes (reviewed by...

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Bacterial Peptides Targeting Periodontal Pathogens in Communities

Xie

Lamont

2020

Emerging Therapies in Periodontics

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A streptococcal effector protein that inhibits Porphyromonas gingivalis biofilm development

Cited by 51 publications

References 48 publications

Arginine deiminase inhibits Porphyromonas gingivalis surface attachment

Arginine deiminase inhibits Porphyromonas gingivalis surface attachment

The nucleoid-associated protein HUβ affects global gene expression in Porphyromonas gingivalis

Bacterial Peptides Targeting Periodontal Pathogens in Communities

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