Extracellular actin is a receptor for Mycoplasma hyopneumoniae

Raymond, BBA; Madhkoor, R; Schleicher, I; Uphoff, CC; Turnbull, L; Whitchurch, CB; Rohde, M; Padula, MP; Djordjevic, SP

Extracellular actin is a receptor for Mycoplasma hyopneumoniae

Raymond, BBA

Madhkoor, R Schleicher, I Uphoff, CC Turnbull, L

Whitchurch, CB

Rohde, M Padula, MP

Djordjevic, SP

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Publication Type:: Journal Article
Citation:: Frontiers in Cellular and Infection Microbiology, 2018, 8 (FEB)
Issue Date:: 2018-02-27

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Field	Value	Language
dc.contributor.author	Raymond, BBA https://orcid.org/0000-0003-3610-9289	en_US
dc.contributor.author	Madhkoor, R	en_US
dc.contributor.author	Schleicher, I	en_US
dc.contributor.author	Uphoff, CC	en_US
dc.contributor.author	Turnbull, L https://orcid.org/0000-0002-9255-9033	en_US
dc.contributor.author	Whitchurch, CB https://orcid.org/0000-0003-2296-3791	en_US
dc.contributor.author	Rohde, M	en_US
dc.contributor.author	Padula, MP https://orcid.org/0000-0002-8283-0643	en_US
dc.contributor.author	Djordjevic, SP https://orcid.org/0000-0001-9301-5372	en_US
dc.date.available	2018-02-12	en_US
dc.date.issued	2018-02-27	en_US
dc.identifier.citation	Frontiers in Cellular and Infection Microbiology, 2018, 8 (FEB)	en_US
dc.identifier.uri	http://hdl.handle.net/10453/124063
dc.description.abstract	© 2018 Raymond, Madhkoor, Schleicher, Uphoff, Turnbull, Whitchurch, Rohde, Padula and Djordjevic. Mycoplasma hyopneumoniae, an agriculturally important porcine pathogen, disrupts the mucociliary escalator causing ciliostasis, loss of cilial function, and epithelial cell death within the porcine lung. Losses to swine production due to growth rate retardation and reduced feed conversion efficiency are severe, and antibiotics are used heavily to control mycoplasmal pneumonia. Notably, little is known about the repertoire of host receptors that M. hyopneumoniae targets to facilitate colonization. Here we show, for the first time, that actin exists extracellularly on porcine epithelial monolayers (PK-15) using surface biotinylation and 3D-Structured Illumination Microscopy (3D-SIM), and that M. hyopneumoniae binds to the extracellular β-actin exposed on the surface of these cells. Consistent with this hypothesis we show: (i) monoclonal antibodies that target β-actin significantly block the ability of M. hyopneumoniae to adhere and colonize PK-15 cells; (ii) microtiter plate binding assays show that M. hyopneumoniae cells bind to monomeric G-actin in a dose dependent manner; (iii) more than 100 M. hyopneumoniae proteins were recovered from affinity-chromatography experiments using immobilized actin as bait; and (iv) biotinylated monomeric actin binds directly to M. hyopneumoniae proteins in ligand blotting studies. Specifically, we show that the P97 cilium adhesin possesses at least two distinct actin-binding regions, and binds monomeric actin with nanomolar affinity. Taken together, these observations suggest that actin may be an important receptor for M. hyopneumoniae within the swine lung and will aid in the future development of intervention strategies against this devastating pathogen. Furthermore, our observations have wider implications for extracellular actin as an important bacterial receptor.	en_US
dc.relation.ispartof	Frontiers in Cellular and Infection Microbiology	en_US
dc.relation.isbasedon	10.3389/fcimb.2018.00054	en_US
dc.subject.mesh	Lung	en_US
dc.subject.mesh	Cell Line	en_US
dc.subject.mesh	Cilia	en_US
dc.subject.mesh	Epithelial Cells	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Swine	en_US
dc.subject.mesh	Mycoplasma hyopneumoniae	en_US
dc.subject.mesh	Pneumonia of Swine, Mycoplasmal	en_US
dc.subject.mesh	Actins	en_US
dc.subject.mesh	Avidin	en_US
dc.subject.mesh	Adhesins, Bacterial	en_US
dc.subject.mesh	Membrane Proteins	en_US
dc.subject.mesh	Antibodies, Monoclonal	en_US
dc.subject.mesh	Biotinylation	en_US
dc.subject.mesh	Protein Binding	en_US
dc.subject.mesh	Host-Pathogen Interactions	en_US
dc.title	Extracellular actin is a receptor for Mycoplasma hyopneumoniae	en_US
dc.type	Journal Article
utslib.citation.volume	FEB	en_US
utslib.citation.volume	8	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	0601 Biochemistry and Cell Biology	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	open_access
pubs.issue	FEB	en_US
pubs.publication-status	Published	en_US
pubs.volume	8	en_US

Abstract:

© 2018 Raymond, Madhkoor, Schleicher, Uphoff, Turnbull, Whitchurch, Rohde, Padula and Djordjevic. Mycoplasma hyopneumoniae, an agriculturally important porcine pathogen, disrupts the mucociliary escalator causing ciliostasis, loss of cilial function, and epithelial cell death within the porcine lung. Losses to swine production due to growth rate retardation and reduced feed conversion efficiency are severe, and antibiotics are used heavily to control mycoplasmal pneumonia. Notably, little is known about the repertoire of host receptors that M. hyopneumoniae targets to facilitate colonization. Here we show, for the first time, that actin exists extracellularly on porcine epithelial monolayers (PK-15) using surface biotinylation and 3D-Structured Illumination Microscopy (3D-SIM), and that M. hyopneumoniae binds to the extracellular β-actin exposed on the surface of these cells. Consistent with this hypothesis we show: (i) monoclonal antibodies that target β-actin significantly block the ability of M. hyopneumoniae to adhere and colonize PK-15 cells; (ii) microtiter plate binding assays show that M. hyopneumoniae cells bind to monomeric G-actin in a dose dependent manner; (iii) more than 100 M. hyopneumoniae proteins were recovered from affinity-chromatography experiments using immobilized actin as bait; and (iv) biotinylated monomeric actin binds directly to M. hyopneumoniae proteins in ligand blotting studies. Specifically, we show that the P97 cilium adhesin possesses at least two distinct actin-binding regions, and binds monomeric actin with nanomolar affinity. Taken together, these observations suggest that actin may be an important receptor for M. hyopneumoniae within the swine lung and will aid in the future development of intervention strategies against this devastating pathogen. Furthermore, our observations have wider implications for extracellular actin as an important bacterial receptor.

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http://hdl.handle.net/10453/124063