Global Gene Expression in
            <i>Staphylococcus aureus</i>
            Biofilms

Beenken, Karen E.; Dunman, Paul M.; McAleese, Fionnuala; Macapagal, Daphne; Murphy, Ellen; Projan, Steven J.; Blevins, Jon S.; Smeltzer, Mark S.

doi:10.1128/jb.186.14.4665-4684.2004

Cited by 537 publications

(544 citation statements)

References 79 publications

(80 reference statements)

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“…This difference was statistically significant (P Ͻ 0.05, Dunn's test), suggesting that KB1050 forms less biofilm in vivo. As reported (15), an isogenic sarA mutant (UAMS-929) was also impaired in its ability to form a biofilm in this model of infection, displaying a 1 log decrease in cfu per catheter relative to UAMS-1 (Fig. 6).…”

Section: The Cida Mutation Affects Biofilm Formation In Other Models Ofmentioning

confidence: 76%

“…Several S. aureus genes have been reported to contribute to its biofilm-forming ability, including agr (3)(4)(5), sarA (5-7), sigB (8), ica (9), rbf (10), tcaR (11), arlRS (12,13), and alsSD (14). Microarray analyses of S. aureus (15,16), Pseudomonas aeruginosa (17,18), Bacillus subtilis (19,20), and Escherichia coli (21) have also illustrated that there are major differences in gene expression between biofilm and planktonic cultures of these organisms. These results underscore the need to study biofilm physiology as a way to better understand how the bacteria exist in vivo.…”

Section: O Ne Of the Primary Causative Agents Of Nosocomial Infectionmentioning

confidence: 99%

“…The effect of the cidA mutation on biofilm development was also assessed in vivo in a previously described murine model of catheter-associated infection (15). After 10 days, the infected catheters of each group of mice were removed and the bacteria recovered from each was quantified by dilution plating and reported as the average cfu per catheter (Fig.…”

Section: The Cida Mutation Affects Biofilm Formation In Other Models Ofmentioning

confidence: 99%

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The cidA murein hydrolase regulator contributes to DNA release and biofilm development in Staphylococcus aureus

Rice

Mann

Endres

et al. 2007

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

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671

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The Staphylococcus aureus cidA and lrgA genes have been shown to affect cell lysis under a variety of conditions during planktonic growth. It is hypothesized that these genes encode holins and antiholins, respectively, and may serve as molecular control elements of bacterial cell lysis. To examine the biological role of cell death and lysis, we studied the impact of the cidA mutation on biofilm development. Interestingly, this mutation had a dramatic impact on biofilm morphology and adherence. The cidA mutant (KB1050) biofilm exhibited a rougher appearance compared with the parental strain (UAMS-1) and was less adherent. Propidium iodide staining revealed that KB1050 accumulated more dead cells within the biofilm population relative to UAMS-1, indicative of reduced cell lysis. In agreement with this finding, quantitative real-time PCR experiments demonstrated the presence of 5-fold less genomic DNA in the KB1050 biofilm relative to UAMS-1. Furthermore, treatment of the UAMS-1 biofilm with DNase I caused extensive cell detachment, whereas similar treatment of the KB1050 biofilm had only a modest effect. These results demonstrate that cidA-controlled cell lysis plays a significant role during biofilm development and that released genomic DNA is an important structural component of S. aureus biofilm.autolysis ͉ extracellular DNA ͉ programmed cell death ͉ holin

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Section: The Cida Mutation Affects Biofilm Formation In Other Models Ofmentioning

confidence: 76%

Section: O Ne Of the Primary Causative Agents Of Nosocomial Infectionmentioning

confidence: 99%

Section: The Cida Mutation Affects Biofilm Formation In Other Models Ofmentioning

confidence: 99%

See 1 more Smart Citation

The cidA murein hydrolase regulator contributes to DNA release and biofilm development in Staphylococcus aureus

Rice

Mann

Endres

et al. 2007

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

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610

671

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“…Although there is substantial evidence showing that arginine catabolism plays a central role in the switch to anaerobic and biofilm growth in many host-associated bacteria, there are few data in regard to arginine availability directing biofilm formation (Beenken et al, 2004; Müsken et al, 2010;Zhu et al, 2010 Zhu et al, , 2007. In agreement with data presented here, arginine has been shown to stimulate biofilm formation of P. aeruginosa (Bernier et al, 2011;Caiazza & O'Toole, 2004).…”

Section: Discussionmentioning

confidence: 99%

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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“…It may be necessary to set up multiple experiments with varying flow rates, especially when comparing S. aureus mutants thought to be impaired in biofilm formation. 12 The relationship between OD and viable cell counts depends on many factors and should be determined empirically for each isolate of S. aureus.…”

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confidence: 99%

Investigation of Biofilm Formation in Clinical Isolates of Staphylococcus aureus

Cassat

Lee

Smeltzer

2007

Methods in Molecular Biology

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SummaryAs with many other bacterial species, the most commonly used method to assess staphylococcal biofilm formation in vitro is the microtiter plate assay. This assay is particularly useful for comparison of multiple strains including large-scale screens of mutant libraries. When such screens are applied to the coagulase-negative staphylococci in general, and Staphylococcus epidermidis in particular, they are relatively straightforward by comparison with microtiter plate assays used to assess biofilm formation in other bacterial species. However, in the case of clinical isolates of Staphylococcus aureus, including methicillin-resistant S. aureus, we have found it necessary to employ specific modifications including precoating of the wells of the microtiter plate with plasma proteins and supplementation of the medium with both salt and glucose. In this chapter, we describe the microtiter plate assay in the specific context of clinical isolates of S. aureus and the use of these modifications. A second in vitro method, which also is generally dependent on coating with plasma proteins and supplementation of the growth medium, is the use of flow cells. In this method, bacteria are allowed to attach to a surface and then monitored with respect to their ability to remain attached to the substrate and differentiate into mature biofilms under the constant pressure of fluid shear force. Although flow cells are not applicable to largescale screens, we have found that they provide a more reproducible and accurate assessment of the capacity of S. aureus clinical isolates to form a biofilm. They also provide a means of analyzing structural differences in biofilm architecture and isolating bacteria and/or spent media for analysis of physiological and metabolic changes associated with the adaptive response to growth in a biofilm. While a primary focus of this chapter is on the use of in vitro assays to assess biofilm formation in clinical isolates of S. aureus, it is important to emphasize two additional considerations. First, it has become increasingly evident that biofilm formation in S. epiderimidis and S. aureus is not equivalent. Additionally, to date, most studies with S. aureus have been done with a very limited number of strains, almost all of which are derived from the NCTC strain designated 8325, and we have found that these strains are not representative of the most relevant clinical isolates. As with the specific elements of our flow cell system, we have written this chapter to reflect our focus on clinical isolates of S. aureus and the specific methods that we have found most reliable in that context. Second, as is often the case, in vitro methods do not necessarily reflect events that occur in vivo. Several in vivo methods to assess biofilm formation have been described, and these generally fall into one of two categories. The first focuses directly on staphylococcal diseases that are generally thought to include a biofilm component (e.g., endocarditis, osteomyelitis, septic arthritis). A discussion of these mod...

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Global Gene Expression in Staphylococcus aureus Biofilms

Cited by 537 publications

References 79 publications

The cidA murein hydrolase regulator contributes to DNA release and biofilm development in Staphylococcus aureus

The cidA murein hydrolase regulator contributes to DNA release and biofilm development in Staphylococcus aureus

Arginine deiminase inhibits Porphyromonas gingivalis surface attachment

Investigation of Biofilm Formation in Clinical Isolates of Staphylococcus aureus

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