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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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In vitro biofilm formation. A. S. aureus SA113 strains adhering to polystyrene microtitre plates were stained with safranin. Four replica are shown. B. Biofilm formation as in (A) but in glass tubes. Three replica are shown.
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fig09: In vitro biofilm formation. A. S. aureus SA113 strains adhering to polystyrene microtitre plates were stained with safranin. Four replica are shown. B. Biofilm formation as in (A) but in glass tubes. Three replica are shown.

Mentions: SA113 strains were also analysed for their capacity to form biofilms during growth in polystyrene microtitre plates (Fig. 9A) or in glass tubes (Fig. 9B). While the wild type clearly formed biofilms on polystyrene the ypfP mutant seemed not to adhere and had completely disappeared from the wells after washing. Biofilm formation was even stronger reduced with the ypfP mutant than with the PIA-deficient ica mutant (Cramton et al., 1999). Wild-type phenotype was restored in the complemented mutant. In glass tubes, only the ica mutant exhibited a defect in biofilm formation. However, the ypfP mutant formed a biofilm on glass confirming the notion that changes in hydrophobicity of either bacteria or biomaterials are crucial for the ability to form biofilms although the exact nature of molecular interactions remain elusive. As the RN4220 wild type showed hardly any biofilm formation the RN4220 was omitted from this experiment. Taken together, our study indicates that the LTA content governs the physicochemical surface properties of S. aureus and enables biofilm formation.


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)

In vitro biofilm formation. A. S. aureus SA113 strains adhering to polystyrene microtitre plates were stained with safranin. Four replica are shown. B. Biofilm formation as in (A) but in glass tubes. Three replica are shown.
© Copyright Policy
Related In: Results  -  Collection

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

fig09: In vitro biofilm formation. A. S. aureus SA113 strains adhering to polystyrene microtitre plates were stained with safranin. Four replica are shown. B. Biofilm formation as in (A) but in glass tubes. Three replica are shown.
Mentions: SA113 strains were also analysed for their capacity to form biofilms during growth in polystyrene microtitre plates (Fig. 9A) or in glass tubes (Fig. 9B). While the wild type clearly formed biofilms on polystyrene the ypfP mutant seemed not to adhere and had completely disappeared from the wells after washing. Biofilm formation was even stronger reduced with the ypfP mutant than with the PIA-deficient ica mutant (Cramton et al., 1999). Wild-type phenotype was restored in the complemented mutant. In glass tubes, only the ica mutant exhibited a defect in biofilm formation. However, the ypfP mutant formed a biofilm on glass confirming the notion that changes in hydrophobicity of either bacteria or biomaterials are crucial for the ability to form biofilms although the exact nature of molecular interactions remain elusive. As the RN4220 wild type showed hardly any biofilm formation the RN4220 was omitted from this experiment. Taken together, our study indicates that the LTA content governs the physicochemical surface properties of S. aureus and enables biofilm formation.

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