Atomic force and super-resolution microscopy support a role for LapA as a cell-surface biofilm adhesin of Pseudomonas fluorescens

Ivanov, IE; Boyd, CD; Newell, PD; Schwartz, ME; Turnbull, L; Johnson, MS; Whitchurch, CB; O'Toole, GA; Camesano, TA

Atomic force and super-resolution microscopy support a role for LapA as a cell-surface biofilm adhesin of Pseudomonas fluorescens

Ivanov, IE Boyd, CD Newell, PD Schwartz, ME Turnbull, L

Johnson, MS Whitchurch, CB

O'Toole, GA Camesano, TA

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Publication Type:: Journal Article
Citation:: Research in Microbiology, 2012, 163 (9-10), pp. 685 - 691
Issue Date:: 2012-11-01

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Field	Value	Language
dc.contributor.author	Ivanov, IE	en_US
dc.contributor.author	Boyd, CD	en_US
dc.contributor.author	Newell, PD	en_US
dc.contributor.author	Schwartz, ME	en_US
dc.contributor.author	Turnbull, L https://orcid.org/0000-0002-9255-9033	en_US
dc.contributor.author	Johnson, MS	en_US
dc.contributor.author	Whitchurch, CB https://orcid.org/0000-0003-2296-3791	en_US
dc.contributor.author	O'Toole, GA	en_US
dc.contributor.author	Camesano, TA	en_US
dc.date.available	2012-09-27	en_US
dc.date.issued	2012-11-01	en_US
dc.identifier.citation	Research in Microbiology, 2012, 163 (9-10), pp. 685 - 691	en_US
dc.identifier.issn	0923-2508	en_US
dc.identifier.uri	http://hdl.handle.net/10453/22177
dc.description.abstract	Pseudomonas fluorescence Pf0-1 requires the large repeat protein LapA for stable surface attachment. This study presents direct evidence that LapA is a cell-surface-localized adhesin. Atomic force microscopy (AFM) revealed a significant 2-fold reduction in adhesion force for mutants lacking the LapA protein on the cell surface compared to the wild-type strain. Deletion of lapG, a gene encoding a periplasmic cysteine protease that functions to release LapA from the cell surface, resulted in a 2-fold increase in the force of adhesion. Three-dimensional structured illumination microscopy (3D-SIM) revealed the presence of the LapA protein on the cell surface, consistent with its role as an adhesin. The protein is only visualized in the cytoplasm for a mutant of the ABC transporter responsible for translocating LapA to the cell surface. Together, these data highlight the power of combining the use of AFM and 3D-SIM with genetic studies to demonstrate that LapA, a member of a large group of RTX-like repeat proteins, is a cell-surface adhesin. © 2012 Institut Pasteur.	en_US
dc.relation.ispartof	Research in Microbiology	en_US
dc.relation.isbasedon	10.1016/j.resmic.2012.10.001	en_US
dc.subject.classification	Microbiology	en_US
dc.subject.mesh	Biofilms	en_US
dc.subject.mesh	Pseudomonas fluorescens	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	Adhesins, Bacterial	en_US
dc.subject.mesh	Lectins	en_US
dc.subject.mesh	Membrane Proteins	en_US
dc.subject.mesh	Microscopy	en_US
dc.subject.mesh	Gene Deletion	en_US
dc.title	Atomic force and super-resolution microscopy support a role for LapA as a cell-surface biofilm adhesin of Pseudomonas fluorescens	en_US
dc.type	Journal Article
utslib.citation.volume	9-10	en_US
utslib.citation.volume	163	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	1108 Medical Microbiology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	closed_access
pubs.issue	9-10	en_US
pubs.publication-status	Published	en_US
pubs.volume	163	en_US

Abstract:

Pseudomonas fluorescence Pf0-1 requires the large repeat protein LapA for stable surface attachment. This study presents direct evidence that LapA is a cell-surface-localized adhesin. Atomic force microscopy (AFM) revealed a significant 2-fold reduction in adhesion force for mutants lacking the LapA protein on the cell surface compared to the wild-type strain. Deletion of lapG, a gene encoding a periplasmic cysteine protease that functions to release LapA from the cell surface, resulted in a 2-fold increase in the force of adhesion. Three-dimensional structured illumination microscopy (3D-SIM) revealed the presence of the LapA protein on the cell surface, consistent with its role as an adhesin. The protein is only visualized in the cytoplasm for a mutant of the ABC transporter responsible for translocating LapA to the cell surface. Together, these data highlight the power of combining the use of AFM and 3D-SIM with genetic studies to demonstrate that LapA, a member of a large group of RTX-like repeat proteins, is a cell-surface adhesin. © 2012 Institut Pasteur.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/22177