Identification of a Dichelobacter nodosus ferric uptake regulator and determination of its regulatory targets

Parker, D; Kennan, RM; Myers, GS; Paulsen, IT; Rood, JI

Identification of a Dichelobacter nodosus ferric uptake regulator and determination of its regulatory targets

Parker, D Kennan, RM Myers, GS

Paulsen, IT Rood, JI

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Publication Type:: Journal Article
Citation:: Journal of Bacteriology, 2005, 187 (1), pp. 366 - 375
Issue Date:: 2005-01-01

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Field	Value	Language
dc.contributor.author	Parker, D	en_US
dc.contributor.author	Kennan, RM	en_US
dc.contributor.author	Myers, GS https://orcid.org/0000-0002-4756-4810	en_US
dc.contributor.author	Paulsen, IT	en_US
dc.contributor.author	Rood, JI	en_US
dc.date.issued	2005-01-01	en_US
dc.identifier.citation	Journal of Bacteriology, 2005, 187 (1), pp. 366 - 375	en_US
dc.identifier.issn	0021-9193	en_US
dc.identifier.uri	http://hdl.handle.net/10453/39762
dc.description.abstract	The expression of iron regulated genes in bacteria is typically controlled by the ferric uptake regulator (Fur) protein, a global transcriptional repressor that regulates functions as diverse as iron acquisition, oxidative stress, and virulence. We have identified afur homologue in Dichelobacter nodosus, the causative agent of ovine footrot, and shown that it complements an Escherichia colifur mutant. Homology modeling of the D. nodosus Fur protein with the recently solved crystal structure of Fur from Pseudomonas aeruginosa indicated extensive structural conservation. As Southern hybridization analysis of different clinical isolates of D. nodosus indicated that the fur gene was present in all of these strains, the fur gene was insertionally inactivated to determine its functional role. Analysis of these mutants by various techniques did not indicate any significant differences in the expression of known virulence genes or in iron-dependent growth. However, we determined several Fur regulatory targets by two-dimensional gel electrophoresis coupled with mass spectrometry. Analysis of proteins from cytoplasmic, membrane, and extracellular fractions revealed numerous differentially expressed proteins. The transcriptional basis of these differences was analyzed by using quantitative reverse transcriptase PCR. Proteins with increased expression in the fur mutant were homologues of the periplasmic iron binding protein YfeA and a cobalt chelatase, CbiK. Down-regulated proteins included a putative manganese superoxide dismutase and ornithine decarboxylase. Based on these data, it is suggested that in D. nodosus the Fur protein functions as a regulator of iron and oxidative metabolism.	en_US
dc.relation.ispartof	Journal of Bacteriology	en_US
dc.relation.isbasedon	10.1128/JB.187.1.366-375.2005	en_US
dc.subject.classification	Microbiology	en_US
dc.subject.mesh	Dichelobacter nodosus	en_US
dc.subject.mesh	Iron	en_US
dc.subject.mesh	Superoxide Dismutase	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	Amino Acid Sequence	en_US
dc.subject.mesh	Cloning, Molecular	en_US
dc.subject.mesh	Molecular Sequence Data	en_US
dc.subject.mesh	Proteomics	en_US
dc.subject.mesh	Repressor Proteins	en_US
dc.title	Identification of a Dichelobacter nodosus ferric uptake regulator and determination of its regulatory targets	en_US
dc.type	Journal Article
utslib.citation.volume	1	en_US
utslib.citation.volume	187	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	07 Agricultural and Veterinary Sciences	en_US
utslib.for	11 Medical and Health Sciences	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	1	en_US
pubs.publication-status	Published	en_US
pubs.volume	187	en_US

Abstract:

The expression of iron regulated genes in bacteria is typically controlled by the ferric uptake regulator (Fur) protein, a global transcriptional repressor that regulates functions as diverse as iron acquisition, oxidative stress, and virulence. We have identified afur homologue in Dichelobacter nodosus, the causative agent of ovine footrot, and shown that it complements an Escherichia colifur mutant. Homology modeling of the D. nodosus Fur protein with the recently solved crystal structure of Fur from Pseudomonas aeruginosa indicated extensive structural conservation. As Southern hybridization analysis of different clinical isolates of D. nodosus indicated that the fur gene was present in all of these strains, the fur gene was insertionally inactivated to determine its functional role. Analysis of these mutants by various techniques did not indicate any significant differences in the expression of known virulence genes or in iron-dependent growth. However, we determined several Fur regulatory targets by two-dimensional gel electrophoresis coupled with mass spectrometry. Analysis of proteins from cytoplasmic, membrane, and extracellular fractions revealed numerous differentially expressed proteins. The transcriptional basis of these differences was analyzed by using quantitative reverse transcriptase PCR. Proteins with increased expression in the fur mutant were homologues of the periplasmic iron binding protein YfeA and a cobalt chelatase, CbiK. Down-regulated proteins included a putative manganese superoxide dismutase and ornithine decarboxylase. Based on these data, it is suggested that in D. nodosus the Fur protein functions as a regulator of iron and oxidative metabolism.

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http://hdl.handle.net/10453/39762