A global genomic approach uncovers novel components for twitching motility-mediated biofilm expansion in Pseudomonas aeruginosa

Nolan, LM; Whitchurch, CB; Barquist, L; Katrib, M; Boinett, CJ; Mayho, M; Goulding, D; Charles, IG; Filloux, A; Parkhill, J; Cain, AK

A global genomic approach uncovers novel components for twitching motility-mediated biofilm expansion in Pseudomonas aeruginosa

Nolan, LM Whitchurch, CB

Barquist, L Katrib, M Boinett, CJ Mayho, M Goulding, D Charles, IG Filloux, A Parkhill, J Cain, AK

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Publication Type:: Journal Article
Citation:: Microbial genomics, 2018, 4 (11)
Issue Date:: 2018-11-01

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Field	Value	Language
dc.contributor.author	Nolan, LM	en_US
dc.contributor.author	Whitchurch, CB https://orcid.org/0000-0003-2296-3791	en_US
dc.contributor.author	Barquist, L	en_US
dc.contributor.author	Katrib, M	en_US
dc.contributor.author	Boinett, CJ	en_US
dc.contributor.author	Mayho, M	en_US
dc.contributor.author	Goulding, D	en_US
dc.contributor.author	Charles, IG	en_US
dc.contributor.author	Filloux, A	en_US
dc.contributor.author	Parkhill, J	en_US
dc.contributor.author	Cain, AK	en_US
dc.date.issued	2018-11-01	en_US
dc.identifier.citation	Microbial genomics, 2018, 4 (11)	en_US
dc.identifier.uri	http://hdl.handle.net/10453/129386
dc.description.abstract	Pseudomonas aeruginosa is an extremely successful pathogen able to cause both acute and chronic infections in a range of hosts, utilizing a diverse arsenal of cell-associated and secreted virulence factors. A major cell-associated virulence factor, the Type IV pilus (T4P), is required for epithelial cell adherence and mediates a form of surface translocation termed twitching motility, which is necessary to establish a mature biofilm and actively expand these biofilms. P. aeruginosa twitching motility-mediated biofilm expansion is a coordinated, multicellular behaviour, allowing cells to rapidly colonize surfaces, including implanted medical devices. Although at least 44 proteins are known to be involved in the biogenesis, assembly and regulation of the T4P, with additional regulatory components and pathways implicated, it is unclear how these components and pathways interact to control these processes. In the current study, we used a global genomics-based random-mutagenesis technique, transposon directed insertion-site sequencing (TraDIS), coupled with a physical segregation approach, to identify all genes implicated in twitching motility-mediated biofilm expansion in P. aeruginosa. Our approach allowed identification of both known and novel genes, providing new insight into the complex molecular network that regulates this process in P. aeruginosa. Additionally, our data suggest that the flagellum-associated gene products have a differential effect on twitching motility, based on whether components are intra- or extracellular. Overall the success of our TraDIS approach supports the use of this global genomic technique for investigating virulence genes in bacterial pathogens.	en_US
dc.relation.ispartof	Microbial genomics	en_US
dc.relation.isbasedon	10.1099/mgen.0.000229	en_US
dc.subject.mesh	Fimbriae, Bacterial	en_US
dc.subject.mesh	Flagella	en_US
dc.subject.mesh	Biofilms	en_US
dc.subject.mesh	Pseudomonas aeruginosa	en_US
dc.subject.mesh	Virulence Factors	en_US
dc.subject.mesh	Microscopy, Electron, Transmission	en_US
dc.subject.mesh	Genomics	en_US
dc.subject.mesh	Mutagenesis	en_US
dc.subject.mesh	Locomotion	en_US
dc.subject.mesh	Genes, Bacterial	en_US
dc.title	A global genomic approach uncovers novel components for twitching motility-mediated biofilm expansion in Pseudomonas aeruginosa	en_US
dc.type	Journal Article
utslib.citation.volume	11	en_US
utslib.citation.volume	4	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	0604 Genetics	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/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	open_access
pubs.issue	11	en_US
pubs.publication-status	Published	en_US
pubs.volume	4	en_US

Abstract:

Pseudomonas aeruginosa is an extremely successful pathogen able to cause both acute and chronic infections in a range of hosts, utilizing a diverse arsenal of cell-associated and secreted virulence factors. A major cell-associated virulence factor, the Type IV pilus (T4P), is required for epithelial cell adherence and mediates a form of surface translocation termed twitching motility, which is necessary to establish a mature biofilm and actively expand these biofilms. P. aeruginosa twitching motility-mediated biofilm expansion is a coordinated, multicellular behaviour, allowing cells to rapidly colonize surfaces, including implanted medical devices. Although at least 44 proteins are known to be involved in the biogenesis, assembly and regulation of the T4P, with additional regulatory components and pathways implicated, it is unclear how these components and pathways interact to control these processes. In the current study, we used a global genomics-based random-mutagenesis technique, transposon directed insertion-site sequencing (TraDIS), coupled with a physical segregation approach, to identify all genes implicated in twitching motility-mediated biofilm expansion in P. aeruginosa. Our approach allowed identification of both known and novel genes, providing new insight into the complex molecular network that regulates this process in P. aeruginosa. Additionally, our data suggest that the flagellum-associated gene products have a differential effect on twitching motility, based on whether components are intra- or extracellular. Overall the success of our TraDIS approach supports the use of this global genomic technique for investigating virulence genes in bacterial pathogens.

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http://hdl.handle.net/10453/129386