DOI: 10.1128/iai.01143-06. Isolation and Characterization of Biofilm Formation-Defective Mutants of <i>Staphylococcus aureus</i>

Isolation and Characterization of Biofilm Formation-Defective Mutants of Staphylococcus aureus

10.1128/iai.01143-06

Crossref journal-article

American Society for Microbiology

Infection and Immunity (235)

Abstract

ABSTRACT Staphylococcus aureus produces biofilm and this mode of colonization facilitates infections that are often difficult to treat and engender high morbidity and mortality. We have exploited bacteriophage Mu transposition methods to create an insertional mutant library in a highly biofilm-forming S. aureus clinical isolate. Our screen identified 38 insertions in 23 distinct genes together with one intergenic region that significantly reduced biofilm formation. Nineteen insertions were mapped in loci not previously known to affect biofilm in this organism. These include insertions in codY , srrA , mgrA , and fmtA , a putative DEAD-box helicase, two members of the zinc-metallo-β lactamase/β-CASP family, and a hypothetical protein with a GGDEF motif. Fifteen insertions occurred in the icaADBC operon, which produces intercellular adhesion antigen (PIA) and is important for biofilm formation in many strains of S. aureus and Staphylococcus epidermidis . Obtaining a high proportion of independent Em-Mu disruptions in icaADBC demonstrated both the importance of PIA for biofilm formation in this clinical strain and the strong validation of the screening procedure that concomitantly uncovered additional mutants. All non- ica mutants were further analyzed by immunoblotting and biochemical fractionation for perturbation of PIA and wall teichoic acid. PIA levels were diminished in the majority of non- ica insertional mutants. Three mutant strains were chosen and were functionally complemented for restored biofilm formation by transformation with plasmids carrying the cloned wild-type gene under the control of a xylose-inducible promoter. This is a comprehensive collection of biofilm-defective mutants that underscores the multifactorial genetic program underlying the establishment of biofilm in this insidious pathogen.

Bibliography

Tu Quoc, P. H., Genevaux, P., Pajunen, M., Savilahti, H., Georgopoulos, C., Schrenzel, J., & Kelley, W. L. (2007). Isolation and Characterization of Biofilm Formation-Defective Mutants of Staphylococcus aureus. Infection and Immunity, 75(3), 1079â1088.

Authors 7

Patrick H. Tu Quoc (first)
Pierre Genevaux (additional)
Maria Pajunen (additional)
Harri Savilahti (additional)
Costa Georgopoulos (additional)
Jacques Schrenzel (additional)
William L. Kelley (additional)

References 79 Referenced 118

Dates

Type	When
Created	18 years, 8 months ago (Dec. 12, 2006, 5:40 a.m.)
Deposited	3 years, 5 months ago (March 5, 2022, 2:01 a.m.)
Indexed	3 months, 4 weeks ago (May 7, 2025, 9:32 a.m.)
Issued	18 years, 6 months ago (March 1, 2007)
Published	18 years, 6 months ago (March 1, 2007)
Published Print	18 years, 6 months ago (March 1, 2007)

Funders 0

None

BibTeX

@article{Tu_Quoc_2007, title={Isolation and Characterization of Biofilm Formation-Defective Mutants of Staphylococcus aureus}, volume={75}, ISSN={1098-5522}, url={http://dx.doi.org/10.1128/iai.01143-06}, DOI={10.1128/iai.01143-06}, number={3}, journal={Infection and Immunity}, publisher={American Society for Microbiology}, author={Tu Quoc, Patrick H. and Genevaux, Pierre and Pajunen, Maria and Savilahti, Harri and Georgopoulos, Costa and Schrenzel, Jacques and Kelley, William L.}, year={2007}, month=mar, pages={1079–1088} }

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