A Putative P-Type ATPase Required for Virulence and Resistance to Haem Toxicity in Listeria monocytogenes
Publication Date
February 21, 2012
Journal
PLOS ONE
Authors
Heather P. Mc Laughlin, Qiaobin Xiao, Rosemarie B. Rea, Hualiang Pi, et al
Volume
7
Issue
2
Pages
e30928
DOI
https://dx.plos.org/10.1371/journal.pone.0030928
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0030928
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/22363518
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3283593
Europe PMC
http://europepmc.org/abstract/MED/22363518
Web of Science
000302873700015
Scopus
84857379518
Mendeley
http://www.mendeley.com/research/putative-ptype-atpase-required-virulence-resistance-haem-toxicity-listeria-monocytogenes
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Mendeley 2601 May 09:05 UTC

Mendeley | Further Information

{"title"=>"A putative P-type ATPase required for virulence and resistance to haem toxicity in listeria monocytogenes", "type"=>"journal", "authors"=>[{"first_name"=>"Heather P.", "last_name"=>"McLaughlin", "scopus_author_id"=>"37120694700"}, {"first_name"=>"Qiaobin", "last_name"=>"Xiao", "scopus_author_id"=>"15847248700"}, {"first_name"=>"Rosemarie B.", "last_name"=>"Rea", "scopus_author_id"=>"7005728109"}, {"first_name"=>"Hualiang", "last_name"=>"Pi", "scopus_author_id"=>"57188847217"}, {"first_name"=>"Pat G.", "last_name"=>"Casey", "scopus_author_id"=>"7102742190"}, {"first_name"=>"Trevor", "last_name"=>"Darby", "scopus_author_id"=>"55025549500"}, {"first_name"=>"Alain", "last_name"=>"Charbit", "scopus_author_id"=>"7006991297"}, {"first_name"=>"Roy D.", "last_name"=>"Sleator", "scopus_author_id"=>"6602741731"}, {"first_name"=>"Susan A.", "last_name"=>"Joyce", "scopus_author_id"=>"7007076890"}, {"first_name"=>"Richard E.", "last_name"=>"Cowart", "scopus_author_id"=>"7003784274"}, {"first_name"=>"Colin", "last_name"=>"Hill", "scopus_author_id"=>"7403558967"}, {"first_name"=>"Phillip E.", "last_name"=>"Klebba", "scopus_author_id"=>"7004643200"}, {"first_name"=>"Cormac G.M.", "last_name"=>"Gahan", "scopus_author_id"=>"7004069227"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"doi"=>"10.1371/journal.pone.0030928", "issn"=>"19326203", "sgr"=>"84857379518", "scopus"=>"2-s2.0-84857379518", "pui"=>"364320885", "pmid"=>"22363518", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)"}, "id"=>"bbd69d2e-89fd-31a6-84d7-874246f609de", "abstract"=>"Regulation of iron homeostasis in many pathogens is principally mediated by the ferric uptake regulator, Fur. Since acquisition of iron from the host is essential for the intracellular pathogen Listeria monocytogenes, we predicted the existence of Fur-regulated systems that support infection. We examined the contribution of nine Fur-regulated loci to the pathogenicity of L. monocytogenes in a murine model of infection. While mutating the majority of the genes failed to affect virulence, three mutants exhibited a significantly compromised virulence potential. Most striking was the role of the membrane protein we designate FrvA (Fur regulated virulence factor A; encoded by frvA [lmo0641]), which is absolutely required for the systemic phase of infection in mice and also for virulence in an alternative infection model, the Wax Moth Galleria mellonella. Further analysis of the ΔfrvA mutant revealed poor growth in iron deficient media and inhibition of growth by micromolar concentrations of haem or haemoglobin, a phenotype which may contribute to the attenuated growth of this mutant during infection. Uptake studies indicated that the ΔfrvA mutant is unaffected in the uptake of ferric citrate but demonstrates a significant increase in uptake of haem and haemin. The data suggest a potential role for FrvA as a haem exporter that functions, at least in part, to protect the cell against the potential toxicity of free haem.", "link"=>"http://www.mendeley.com/research/putative-ptype-atpase-required-virulence-resistance-haem-toxicity-listeria-monocytogenes", "reader_count"=>26, "reader_count_by_academic_status"=>{"Student > Doctoral Student"=>6, "Researcher"=>3, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>1, "Student > Master"=>2, "Student > Bachelor"=>2, "Professor"=>1}, "reader_count_by_user_role"=>{"Student > Doctoral Student"=>6, "Researcher"=>3, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>1, "Student > Master"=>2, "Student > Bachelor"=>2, "Professor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>20, "Medicine and Dentistry"=>1, "Chemistry"=>1, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>20}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}}, "reader_count_by_country"=>{"Portugal"=>1}, "group_count"=>0}
Scopus 2811 Feb 20:06 UTC

Scopus | Further Information

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Figshare 4102 Apr 19:36 UTC

Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/346186", "https://ndownloader.figshare.com/files/346250", "https://ndownloader.figshare.com/files/346303", "https://ndownloader.figshare.com/files/346364", "https://ndownloader.figshare.com/files/346429"], "description"=>"<div><p>Regulation of iron homeostasis in many pathogens is principally mediated by the ferric uptake regulator, Fur. Since acquisition of iron from the host is essential for the intracellular pathogen <em>Listeria monocytogenes</em>, we predicted the existence of Fur-regulated systems that support infection. We examined the contribution of nine Fur-regulated loci to the pathogenicity of <em>L. monocytogenes</em> in a murine model of infection. While mutating the majority of the genes failed to affect virulence, three mutants exhibited a significantly compromised virulence potential. Most striking was the role of the membrane protein we designate FrvA (<u>F</u>ur <u>r</u>egulated <u>v</u>irulence factor A; encoded by <em>frvA</em> [<em>lmo0641</em>]), which is absolutely required for the systemic phase of infection in mice and also for virulence in an alternative infection model, the Wax Moth <em>Galleria mellonella</em>. Further analysis of the <em>ΔfrvA</em> mutant revealed poor growth in iron deficient media and inhibition of growth by micromolar concentrations of haem or haemoglobin, a phenotype which may contribute to the attenuated growth of this mutant during infection. Uptake studies indicated that the <em>ΔfrvA</em> mutant is unaffected in the uptake of ferric citrate but demonstrates a significant increase in uptake of haem and haemin. The data suggest a potential role for FrvA as a haem exporter that functions, at least in part, to protect the cell against the potential toxicity of free haem.</p> </div>", "links"=>[], "tags"=>["putative", "p-type", "atpase", "virulence", "haem", "toxicity"], "article_id"=>128416, "categories"=>["Cancer", "Microbiology"], "users"=>["Heather P. McLaughlin", "Qiaobin Xiao", "Rosemarie B. Rea", "Hualiang Pi", "Pat G. Casey", "Trevor Darby", "Alain Charbit", "Roy D. Sleator", "Susan A. Joyce", "Richard E. Cowart", "Colin Hill", "Phillip E. Klebba", "Cormac G. M. Gahan"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030928.s001", "https://dx.doi.org/10.1371/journal.pone.0030928.s002", "https://dx.doi.org/10.1371/journal.pone.0030928.s003", "https://dx.doi.org/10.1371/journal.pone.0030928.s004", "https://dx.doi.org/10.1371/journal.pone.0030928.s005"], "stats"=>{"downloads"=>10, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/A_Putative_P_Type_ATPase_Required_for_Virulence_and_Resistance_to_Haem_Toxicity_in_Listeria_monocytogenes_/128416", "title"=>"A Putative P-Type ATPase Required for Virulence and Resistance to Haem Toxicity in <em>Listeria monocytogenes</em>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-02-21 02:20:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/678589"], "description"=>"<p>The rates and extent of bacterial growth (A: EGD-e; C: Δ<i>lmo0641</i>; E:Δ<i>lmo0641</i>/pPL2<i>lmo0641</i>) were determined in iron-restricted MOPS-L media supplemented with Hb (panels <b>A</b>–<b>C</b>; open, gray and black symbols represent addition of 0.0, 0.02 and 2 uM Hb, respectively) or Hn (<b>D</b>, <b>E</b>; open, gray and black symbols represent addition of 0.0, 0.2 and 2 uM Hn, respectively), and in BHI broth (<b>F</b>). The bacteria were cultured in BHI broth overnight. In <b>A–E</b> they were then subcultured in MOPS-L to stationary phase, and at t = 0 subcultured again at 1% into MOPS-L containing different concentrations of Hb or Hn. In <b>F</b>, at t = 0 they were subcultured into BHI broth. The flasks were shaken at 37°C and absorbance at 600 nm (initially close to zero for all cultures) was monitored for 12–26 h (note different scales). Because of the slow growth of <i>L. monocytogenes</i> in iron-restricted minimal media, this graphic representation focuses on the comparison of the mutant strains at later times in the growth cycle.</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>349069, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Heather P. McLaughlin", "Qiaobin Xiao", "Rosemarie B. Rea", "Hualiang Pi", "Pat G. Casey", "Trevor Darby", "Alain Charbit", "Roy D. Sleator", "Susan A. Joyce", "Richard E. Cowart", "Colin Hill", "Phillip E. Klebba", "Cormac G. M. Gahan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030928.g004", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bacterial_growth_/349069", "title"=>"Bacterial growth.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-21 02:31:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/678456"], "description"=>"<p>Uptake affinity (K<sub>m</sub> in nM) and velocity (V<sub>max</sub> in pMol per 10<sup>9</sup> cells per minute) by which the wild-type (open circles) and Δ<i>0641</i> (Δ<i>frvA</i>) (closed circles) strains transport [<sup>59</sup>Fe]-citrate (A) and [<sup>59</sup>Fe]-Hn (B) were assessed. Overall K<sub>m</sub> and V<sub>max</sub> of [<sup>59</sup>Fe] transport are listed in the tables on right-hand side. Data was plotted using the Enzyme Kinetics algorithm of Grafit 7 (Erithacus Ltd, West Sussex, UK) and represent the mean of independent experiments done in triplicate.</p>", "links"=>[], "tags"=>["binding", "uptake"], "article_id"=>348935, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Heather P. McLaughlin", "Qiaobin Xiao", "Rosemarie B. Rea", "Hualiang Pi", "Pat G. Casey", "Trevor Darby", "Alain Charbit", "Roy D. Sleator", "Susan A. Joyce", "Richard E. Cowart", "Colin Hill", "Phillip E. Klebba", "Cormac G. M. Gahan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030928.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_59_Fe_binding_and_uptake_assays_/348935", "title"=>"<sup>59</sup>Fe binding and uptake assays.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-21 02:28:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/678249"], "description"=>"<p>(A) The classical Fur box is represented as a 19 bp sequence. Recent studies have suggested that a more accurate representation of the Fur box is that of a 7-1-7 motif. The 19 bp sequence was used to search the <i>Listeria monocytogenes</i> EGDe genome sequence (Listilist). (B) Identified sequences were aligned and a graphical display of the results was generated using the web based programme sequence logo (17). (C) Genetic organisation of 29 putative Fur regulated genes (black/gray) at 12 chromosomal loci. All genes are drawn approximately to scale using the <i>L. monocytogenes</i> EGDe genome sequence data. Lmo numbers refer to the National Centre for Biotechnology Information annotation scheme. Fur boxes are represented by black circles. Gray genes indicate those disrupted in EGDe in the course of this study. Lollipops are used to illustrate putative stem loop terminator regions. (D) RT-PCR analysis was used to confirm Fur regulation of all identified genes and to give an indication of the increase in expression levels. Control primers were used to ensure that template cDNAs were of equal concentration. Samples were removed at various cycles of PCR (cycle number in brackets) and visualised on agarose gels. A repeat experiment demonstrated similar results. Results were also verified through real-time PCR analysis. (E) In vivo survival of disruption mutants in Fur-regulated loci in the murine infection model. Mice were injected i.p. with either the wild-type or mutants and the number of bacteria recovered from the spleen was determined three days post-inoculation. Error bars represent the standard deviations from the mean (n = 4). * indicates means are significantly different to the wild-type (P<0.05). ND, not detected.</p>", "links"=>[], "tags"=>["virulence", "fur-regulated"], "article_id"=>348724, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Heather P. McLaughlin", "Qiaobin Xiao", "Rosemarie B. Rea", "Hualiang Pi", "Pat G. Casey", "Trevor Darby", "Alain Charbit", "Roy D. Sleator", "Susan A. Joyce", "Richard E. Cowart", "Colin Hill", "Phillip E. Klebba", "Cormac G. M. Gahan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030928.g001", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Identification_and_role_in_virulence_of_Fur_regulated_gene_systems_/348724", "title"=>"Identification and role in virulence of Fur-regulated gene systems.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-21 02:25:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/678721"], "description"=>"<p>Tests demonstrate the halo of growth surrounding a paper disc embedded with 10 µl aliquots of the test iron compound. Concentrations of compounds are indicated as µM. Fc (ferrichrome) and FcA (ferrichrome A), FxB (ferrioxamine B), Hb (haem/haemoglobin) and Hn (haemin) were tested on BHI agar containing 0.1 mM BP. The experiment was repeated several times with similar results. No differences were seen between mutant strains and the wild-type in these iron nutrition assays.</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>349200, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Heather P. McLaughlin", "Qiaobin Xiao", "Rosemarie B. Rea", "Hualiang Pi", "Pat G. Casey", "Trevor Darby", "Alain Charbit", "Roy D. Sleator", "Susan A. Joyce", "Richard E. Cowart", "Colin Hill", "Phillip E. Klebba", "Cormac G. M. Gahan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030928.g005", "stats"=>{"downloads"=>4, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Nutrition_tests_/349200", "title"=>"Nutrition tests.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-21 02:33:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/678378"], "description"=>"<p>(A) CFUs of the Δ<i>frvA</i> and Δ<i>frvA</i> complemented strain enumerated from livers and spleens three days post infection. Error bars represent standard error of the mean and asterisks represent P<0.001 by the Student's t-test when compared to the wild-type and complement strains. (B) Pathogenesis of strains in the <i>Galleria mellonella</i> model of infection. Dotted line and cross indicates LT-50 (time in which 50% of insects had perished). (C) Pathogenesis of the Δ<i>lmo0642</i> mutant in the murine model of infection. Strains were inoculated into mice by the ip route and numbers were enumerated in the spleens at day three post-infection. Student t-test did not detect a significant difference between the wild-type and mutant strain.</p>", "links"=>[], "tags"=>["strains", "murine", "wax", "moth", "models"], "article_id"=>348857, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Heather P. McLaughlin", "Qiaobin Xiao", "Rosemarie B. Rea", "Hualiang Pi", "Pat G. Casey", "Trevor Darby", "Alain Charbit", "Roy D. Sleator", "Susan A. Joyce", "Richard E. Cowart", "Colin Hill", "Phillip E. Klebba", "Cormac G. M. Gahan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030928.g002", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pathogenesis_of_strains_in_murine_and_Wax_Moth_models_of_infection_/348857", "title"=>"Pathogenesis of strains in murine and Wax Moth models of infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-21 02:27:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/678887"], "description"=>"<p>Cm<sup>R</sup>, chloramphenicol resistant.</p>", "links"=>[], "tags"=>["strains", "plasmids"], "article_id"=>349367, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Heather P. McLaughlin", "Qiaobin Xiao", "Rosemarie B. Rea", "Hualiang Pi", "Pat G. Casey", "Trevor Darby", "Alain Charbit", "Roy D. Sleator", "Susan A. Joyce", "Richard E. Cowart", "Colin Hill", "Phillip E. Klebba", "Cormac G. M. Gahan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030928.t001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bacterial_strains_and_plasmids_used_in_this_study_/349367", "title"=>"Bacterial strains and plasmids used in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-02-21 02:36:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/678830"], "description"=>"<p>Induction of <i>lmo0641</i> (<i>frvA</i>) transcription in Δ<i>fur</i> compared to the wild-type (black bar) and induction of gene transcriptions in Δ<i>frvA</i> compared to the wild-type (gray bars) in BHI. Up-regulated genes are represented by bars above the x-axis and the down-regulated gene (<i>fur</i>) is represented by the bar below the axis. Asterisks represent Fur-regulated genes. Error bars represent the mean ± SD of the relative change in gene expression of independent duplicate samples.</p>", "links"=>[], "tags"=>["Real-time"], "article_id"=>349311, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Heather P. McLaughlin", "Qiaobin Xiao", "Rosemarie B. Rea", "Hualiang Pi", "Pat G. Casey", "Trevor Darby", "Alain Charbit", "Roy D. Sleator", "Susan A. Joyce", "Richard E. Cowart", "Colin Hill", "Phillip E. Klebba", "Cormac G. M. Gahan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030928.g006", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Quantitative_real_time_PCR_/349311", "title"=>"Quantitative real-time PCR.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-21 02:35:11"}
PMC Usage Stats 119617 Feb 17:11 UTC

PMC Usage Stats | Further Information

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Relative Metric 29265312 Apr 19:00 UTC

Relative Metric

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