Expression of Fap amyloids in Pseudomonas aeruginosa, P. fluorescens, and P. putida results in aggregation and increased biofilm formation
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Expression of Fap amyloids in Pseudomonas aeruginosa, P. fluorescens, and P. putida results in aggregation and increased biofilm formation. / Dueholm, Morten Simonsen; Søndergaard, Mads; Nilsson, Martin; Christiansen, Gunna; Stensballe, Allan; Overgaard, Michael Toft; Givskov, Michael; Tolker-Nielsen, Tim; Otzen, Daniel; Nielsen, Per H.
In: MicrobiologyOpen, Vol. 2, 18.03.2013, p. 365-382.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Expression of Fap amyloids in Pseudomonas aeruginosa, P. fluorescens, and P. putida results in aggregation and increased biofilm formation
AU - Dueholm, Morten Simonsen
AU - Søndergaard, Mads
AU - Nilsson, Martin
AU - Christiansen, Gunna
AU - Stensballe, Allan
AU - Overgaard, Michael Toft
AU - Givskov, Michael
AU - Tolker-Nielsen, Tim
AU - Otzen, Daniel
AU - Nielsen, Per H
N1 - © 2013 The Authors. MicrobiologyOpen published by Blackwell Publishing Ltd.
PY - 2013/3/18
Y1 - 2013/3/18
N2 - The fap operon, encoding functional amyloids in Pseudomonas (Fap), is present in most pseudomonads, but so far the expression and importance for biofilm formation has only been investigated for P. fluorescens strain UK4. In this study, we demonstrate the capacity of P. aeruginosa PAO1, P. fluorescens Pf-5, and P. putida F1 to express Fap fibrils, and investigated the effect of Fap expression on aggregation and biofilm formation. The fap operon in all three Pseudomonas species conferred the ability to express Fap fibrils as shown using a recombinant approach. This Fap overexpression consistently resulted in highly aggregative phenotypes and in increased biofilm formation. Detailed biophysical investigations of purified fibrils confirmed FapC as the main fibril monomer and supported the role of FapB as a minor, nucleating constituent as also indicated by bioinformatic analysis. Bioinformatics analysis suggested FapF and FapD as a potential β-barrel membrane pore and protease, respectively. Manipulation of the fap operon showed that FapA affects monomer composition of the final amyloid fibril, and that FapB is an amyloid protein, probably a nucleator for FapC polymerization. Our study highlights the fap operon as a molecular machine for functional amyloid formation.
AB - The fap operon, encoding functional amyloids in Pseudomonas (Fap), is present in most pseudomonads, but so far the expression and importance for biofilm formation has only been investigated for P. fluorescens strain UK4. In this study, we demonstrate the capacity of P. aeruginosa PAO1, P. fluorescens Pf-5, and P. putida F1 to express Fap fibrils, and investigated the effect of Fap expression on aggregation and biofilm formation. The fap operon in all three Pseudomonas species conferred the ability to express Fap fibrils as shown using a recombinant approach. This Fap overexpression consistently resulted in highly aggregative phenotypes and in increased biofilm formation. Detailed biophysical investigations of purified fibrils confirmed FapC as the main fibril monomer and supported the role of FapB as a minor, nucleating constituent as also indicated by bioinformatic analysis. Bioinformatics analysis suggested FapF and FapD as a potential β-barrel membrane pore and protease, respectively. Manipulation of the fap operon showed that FapA affects monomer composition of the final amyloid fibril, and that FapB is an amyloid protein, probably a nucleator for FapC polymerization. Our study highlights the fap operon as a molecular machine for functional amyloid formation.
U2 - 10.1002/mbo3.81
DO - 10.1002/mbo3.81
M3 - Journal article
C2 - 23504942
VL - 2
SP - 365
EP - 382
JO - MicrobiologyOpen
JF - MicrobiologyOpen
SN - 2045-8827
ER -
ID: 45954660