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A Staphylococcus aureus ypfP mutant with strongly reduced lipoteichoic acid (LTA) content: LTA governs bacterial surface properties and autolysin activity.

Fedtke I, Mader D, Kohler T, Moll H, Nicholson G, Biswas R, Henseler K, Götz F, Zähringer U, Peschel A - Mol. Microbiol. (2007)

Bottom Line: The ypfP gene responsible for biosynthesis of a glycolipid found in LTA was deleted in Staphylococcus aureus SA113, causing 87% reduction of the LTA content.However, the autolytic activity of the mutant was strongly reduced demonstrating a role of LTA in controlling autolysin activity.Moreover, the hydrophobicity of the LTA mutant was altered and its ability to form biofilms on plastic was completely abrogated indicating a profound impact of LTA on physicochemical properties of bacterial surfaces.

View Article: PubMed Central - PubMed

Affiliation: Cellular and Molecular Microbiology Division, University of Tübingen, Department of Medical Microbiology and Hygiene, 72076 Tübingen, Germany.

ABSTRACT
Many Gram-positive bacteria produce lipoteichoic acid (LTA) polymers whose physiological roles have remained a matter of debate because of the lack of LTA-deficient mutants. The ypfP gene responsible for biosynthesis of a glycolipid found in LTA was deleted in Staphylococcus aureus SA113, causing 87% reduction of the LTA content. Mass spectrometry and nuclear magnetic resonance spectroscopy revealed that the mutant LTA contained a diacylglycerol anchor instead of the glycolipid, whereas the remaining part was similar to the wild-type polymer except that it was shorter. The LTA mutant strain revealed no major changes in patterns of cell wall proteins or autolytic enzymes compared with the parental strain indicating that LTA may be less important in S. aureus protein attachment than previously thought. However, the autolytic activity of the mutant was strongly reduced demonstrating a role of LTA in controlling autolysin activity. Moreover, the hydrophobicity of the LTA mutant was altered and its ability to form biofilms on plastic was completely abrogated indicating a profound impact of LTA on physicochemical properties of bacterial surfaces. We propose to consider LTA and its biosynthetic enzymes as targets for new antibiofilm strategies.

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Detection of S. aureus SA113 glycolipids by TLC. Glycolipids were visualized on TLC plates with α-naphthol. The spot missing in the S. aureus ypfP mutant was identified as DGlcDAG.
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fig01: Detection of S. aureus SA113 glycolipids by TLC. Glycolipids were visualized on TLC plates with α-naphthol. The spot missing in the S. aureus ypfP mutant was identified as DGlcDAG.

Mentions: The ypfP gene previously shown to be essential for DGlcDAG biosynthesis (Kiriukhin et al., 2001) was inactivated in S. aureus SA113 by gene replacement. Polar lipids of the resulting mutant were analysed by thin-layer chromatography (TLC) and found to lack a lipid spot present in the wild type. The lipid in question was stainable with α-naphthol, which is indicative of glycolipids (Fig. 1). Complementation of the mutant with a wild-type copy of the deleted gene restored glycolipid production (Fig. 1). The lipid lacking in the mutant was analysed by gas chromatography coupled to mass spectrometry (GC-MS) and found to contain glucose, glycerol (Gro) and fatty acids (predominantly branched C15 acyl chains; data not shown), which is in accordance with the expected constituents of DGlcDAG.


A Staphylococcus aureus ypfP mutant with strongly reduced lipoteichoic acid (LTA) content: LTA governs bacterial surface properties and autolysin activity.

Fedtke I, Mader D, Kohler T, Moll H, Nicholson G, Biswas R, Henseler K, Götz F, Zähringer U, Peschel A - Mol. Microbiol. (2007)

Detection of S. aureus SA113 glycolipids by TLC. Glycolipids were visualized on TLC plates with α-naphthol. The spot missing in the S. aureus ypfP mutant was identified as DGlcDAG.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2169524&req=5

fig01: Detection of S. aureus SA113 glycolipids by TLC. Glycolipids were visualized on TLC plates with α-naphthol. The spot missing in the S. aureus ypfP mutant was identified as DGlcDAG.
Mentions: The ypfP gene previously shown to be essential for DGlcDAG biosynthesis (Kiriukhin et al., 2001) was inactivated in S. aureus SA113 by gene replacement. Polar lipids of the resulting mutant were analysed by thin-layer chromatography (TLC) and found to lack a lipid spot present in the wild type. The lipid in question was stainable with α-naphthol, which is indicative of glycolipids (Fig. 1). Complementation of the mutant with a wild-type copy of the deleted gene restored glycolipid production (Fig. 1). The lipid lacking in the mutant was analysed by gas chromatography coupled to mass spectrometry (GC-MS) and found to contain glucose, glycerol (Gro) and fatty acids (predominantly branched C15 acyl chains; data not shown), which is in accordance with the expected constituents of DGlcDAG.

Bottom Line: The ypfP gene responsible for biosynthesis of a glycolipid found in LTA was deleted in Staphylococcus aureus SA113, causing 87% reduction of the LTA content.However, the autolytic activity of the mutant was strongly reduced demonstrating a role of LTA in controlling autolysin activity.Moreover, the hydrophobicity of the LTA mutant was altered and its ability to form biofilms on plastic was completely abrogated indicating a profound impact of LTA on physicochemical properties of bacterial surfaces.

View Article: PubMed Central - PubMed

Affiliation: Cellular and Molecular Microbiology Division, University of Tübingen, Department of Medical Microbiology and Hygiene, 72076 Tübingen, Germany.

ABSTRACT
Many Gram-positive bacteria produce lipoteichoic acid (LTA) polymers whose physiological roles have remained a matter of debate because of the lack of LTA-deficient mutants. The ypfP gene responsible for biosynthesis of a glycolipid found in LTA was deleted in Staphylococcus aureus SA113, causing 87% reduction of the LTA content. Mass spectrometry and nuclear magnetic resonance spectroscopy revealed that the mutant LTA contained a diacylglycerol anchor instead of the glycolipid, whereas the remaining part was similar to the wild-type polymer except that it was shorter. The LTA mutant strain revealed no major changes in patterns of cell wall proteins or autolytic enzymes compared with the parental strain indicating that LTA may be less important in S. aureus protein attachment than previously thought. However, the autolytic activity of the mutant was strongly reduced demonstrating a role of LTA in controlling autolysin activity. Moreover, the hydrophobicity of the LTA mutant was altered and its ability to form biofilms on plastic was completely abrogated indicating a profound impact of LTA on physicochemical properties of bacterial surfaces. We propose to consider LTA and its biosynthetic enzymes as targets for new antibiofilm strategies.

Show MeSH
Related in: MedlinePlus