Identification of flgZ as a Flagellar Gene Encoding a PilZ Domain Protein That Regulates Swimming Motility and Biofilm Formation in Pseudomonas
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{"title"=>"Identification of flgZ as a flagellar gene encoding a PilZ domain protein that regulates swimming motility and biofilm formation in Pseudomonas", "type"=>"journal", "authors"=>[{"first_name"=>"Francisco", "last_name"=>"Martínez-Granero", "scopus_author_id"=>"8058190500"}, {"first_name"=>"Ana", "last_name"=>"Navazo", "scopus_author_id"=>"24332525700"}, {"first_name"=>"Emma", "last_name"=>"Barahona", "scopus_author_id"=>"24330945400"}, {"first_name"=>"Miguel", "last_name"=>"Redondo-Nieto", "scopus_author_id"=>"7801554314"}, {"first_name"=>"Elena González", "last_name"=>"De Heredia", "scopus_author_id"=>"56054106500"}, {"first_name"=>"Irene", "last_name"=>"Baena", "scopus_author_id"=>"55207564400"}, {"first_name"=>"Irene", "last_name"=>"Martín-Martín", "scopus_author_id"=>"56054780800"}, {"first_name"=>"Rafael", "last_name"=>"Rivilla", "scopus_author_id"=>"6602841580"}, {"first_name"=>"Marta", "last_name"=>"Martín", "scopus_author_id"=>"55482097400"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"24504373", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0087608", "pui"=>"372510324", "isbn"=>"1932-6203 (Electronic)\r1932-6203 (Linking)", "scopus"=>"2-s2.0-84895197830", "sgr"=>"84895197830"}, "id"=>"e31d7a55-0ee5-3376-a8b4-d2e0e1e728bf", "abstract"=>"Diguanylate cyclase and phosphodiesterase enzymatic activities control c-di-GMP levels modulating planktonic versus sessile lifestyle behavior in bacteria. The PilZ domain is described as a sensor of c-di-GMP intracellular levels and the proteins containing a PilZ domain represent the best studied class of c-di-GMP receptors forming part of the c-di-GMP signaling cascade. In P. fluorescens F113 we have found two diguanylate cyclases (WspR, SadC) and one phosphodiesterase (BifA) implicated in regulation of swimming motility and biofilm formation. Here we identify a flgZ gene located in a flagellar operon encoding a protein that contains a PilZ domain. Moreover, we show that FlgZ subcellular localization depends on the c-di-GMP intracellular levels. The overexpression analysis of flgZ in P. fluorescens F113 and P. putida KT2440 backgrounds reveal a participation of FlgZ in Pseudomonas swimming motility regulation. Besides, the epistasis of flgZ over wspR and bifA clearly shows that c-di-GMP intracellular levels produced by the enzymatic activity of the diguanylate cyclase WspR and the phosphodiesterase BifA regulates biofilm formation through FlgZ.", "link"=>"http://www.mendeley.com/research/identification-flgz-flagellar-gene-encoding-pilz-domain-protein-regulates-swimming-motility-biofilm", "reader_count"=>49, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>5, "Researcher"=>15, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>12, "Other"=>1, "Student > Master"=>4, "Student > Bachelor"=>7}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>5, "Researcher"=>15, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>12, "Other"=>1, "Student > Master"=>4, "Student > Bachelor"=>7}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Engineering"=>1, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>6, "Agricultural and Biological Sciences"=>30, "Medicine and Dentistry"=>1, "Physics and Astronomy"=>1, "Chemistry"=>1, "Computer Science"=>1, "Immunology and Microbiology"=>4, "Sports and Recreations"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>4}, "Sports and Recreations"=>{"Sports and Recreations"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>30}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}, "Unspecified"=>{"Unspecified"=>2}, "Environmental Science"=>{"Environmental Science"=>1}}, "reader_count_by_country"=>{"United States"=>1, "China"=>1, "Denmark"=>1, "Spain"=>1}, "group_count"=>5}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1375129"], "description"=>"<p>A. Alignment of the genomic region which contains the <i>flgZ</i> gene in representative strains across the genus <i>Pseudomonas</i>. In all the strains the <i>flgZ</i> gene lies within a flagellar genes region, located downstream the <i>flgMN</i> genes. Other <i>flg</i> and <i>che</i> genes are also conserved in all the strains. B. Domain architecture of the FlgZ protein compared to flagellar brake proteins YcgR from <i>Escherichia coli</i> and DgrA from <i>Caulobacter crescentus</i>. The <i>P. fluorescens</i> F113 contains an N-terminal YcgR domain (PF07317) and a C-Terminal PilZ domain (PF07238). This domain architecture is very similar to that of the YcgR protein. The DgrA protein also contains a PilZ domain. E-value indicates significance of the similarity to the PFAM model domain.</p>", "links"=>[], "tags"=>["biophysics", "Cell motility", "Flagellar motility", "microbiology", "bacteriology", "Bacterial biofilms", "Bacterial pathogens", "Microbial physiology", "Plant microbiology", "Infectious diseases", "Bacterial diseases", "Pseudomonas infections", "conserved", "encodes", "pilz", "homologous"], "article_id"=>925312, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Francisco Martínez-Granero", "Ana Navazo", "Emma Barahona", "Miguel Redondo-Nieto", "Elena González de Heredia", "Irene Baena", "Irene Martín-Martín", "Rafael Rivilla", "Marta Martín"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087608.g001", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_flgZ_is_a_conserved_gene_that_encodes_a_PilZ_protein_homologous_to_YcgR_/925312", "title"=>"<i>flgZ</i> is a conserved gene that encodes a PilZ protein homologous to YcgR.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-04 03:21:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1375137", "https://ndownloader.figshare.com/files/1375138", "https://ndownloader.figshare.com/files/1375139"], "description"=>"<div><p>Diguanylate cyclase and phosphodiesterase enzymatic activities control c-di-GMP levels modulating planktonic versus sessile lifestyle behavior in bacteria. The PilZ domain is described as a sensor of c-di-GMP intracellular levels and the proteins containing a PilZ domain represent the best studied class of c-di-GMP receptors forming part of the c-di-GMP signaling cascade. In <i>P. fluorescens</i> F113 we have found two diguanylate cyclases (WspR, SadC) and one phosphodiesterase (BifA) implicated in regulation of swimming motility and biofilm formation. Here we identify a <i>flgZ</i> gene located in a flagellar operon encoding a protein that contains a PilZ domain. Moreover, we show that FlgZ subcellular localization depends on the c-di-GMP intracellular levels. The overexpression analysis of <i>flgZ</i> in <i>P. fluorescens</i> F113 and <i>P. putida</i> KT2440 backgrounds reveal a participation of FlgZ in Pseudomonas swimming motility regulation. Besides, the epistasis of <i>flgZ</i> over <i>wspR</i> and <i>bifA</i> clearly shows that c-di-GMP intracellular levels produced by the enzymatic activity of the diguanylate cyclase WspR and the phosphodiesterase BifA regulates biofilm formation through FlgZ.</p></div>", "links"=>[], "tags"=>["biophysics", "Cell motility", "Flagellar motility", "microbiology", "bacteriology", "Bacterial biofilms", "Bacterial pathogens", "Microbial physiology", "Plant microbiology", "Infectious diseases", "Bacterial diseases", "Pseudomonas infections", "flagellar", "encoding", "pilz", "regulates", "motility", "biofilm"], "article_id"=>925320, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Francisco Martínez-Granero", "Ana Navazo", "Emma Barahona", "Miguel Redondo-Nieto", "Elena González de Heredia", "Irene Baena", "Irene Martín-Martín", "Rafael Rivilla", "Marta Martín"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087608.s001", "https://dx.doi.org/10.1371/journal.pone.0087608.s002", "https://dx.doi.org/10.1371/journal.pone.0087608.s003"], "stats"=>{"downloads"=>14, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Identification_of_flgZ_as_a_Flagellar_Gene_Encoding_a_PilZ_Domain_Protein_That_Regulates_Swimming_Motility_and_Biofilm_Formation_in_Pseudomonas_/925320", "title"=>"Identification of <i>flgZ</i> as a Flagellar Gene Encoding a PilZ Domain Protein That Regulates Swimming Motility and Biofilm Formation in <i>Pseudomonas</i>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-02-04 03:21:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1375135"], "description"=>"<p>Relative biofilm formation by <i>P. fluorescens</i> F113 mutants affected in c-di-GMP turn-over and sensing. The <i>flgZ</i> mutation suppressed the biofilm forming phenotype of mutants affected in genes encoding the WspR diguanylate cyclase and the BifA phosphodiesterase. Biofilm formation was determined colorimetrically from the crystal violet staining biomass attached to plastic in 96 wells plates, after elimination of the culture grown in these wells in LB medium for 8 h. Averages of three experiments with eight replicates plus standard deviation are shown. Different letters indicate statistically significant differences (p<0.05).</p>", "links"=>[], "tags"=>["biophysics", "Cell motility", "Flagellar motility", "microbiology", "bacteriology", "Bacterial biofilms", "Bacterial pathogens", "Microbial physiology", "Plant microbiology", "Infectious diseases", "Bacterial diseases", "Pseudomonas infections", "modulates", "biofilm"], "article_id"=>925318, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Francisco Martínez-Granero", "Ana Navazo", "Emma Barahona", "Miguel Redondo-Nieto", "Elena González de Heredia", "Irene Baena", "Irene Martín-Martín", "Rafael Rivilla", "Marta Martín"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087608.g006", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_c_di_GMP_modulates_biofilm_formation_through_FlgZ_/925318", "title"=>"c-di-GMP modulates biofilm formation through FlgZ.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-04 03:21:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1375132"], "description"=>"<p>A. Relative swimming motility of <i>P. fluorescens</i> F113 mutants affected in genes encoding diguanylate cyclases (<i>wspR</i> and <i>sadC</i>) and a phosphodiesterase (<i>bifA</i>). Swimming motility was determined by haloes diameter in SA plates with 0.3% agar, 24 hours after inoculation. Averages of three triplicated experiments plus standard deviation are shown. Different letters indicate statistically significant differences (p<0.05). Relative motility of each strain is also shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087608#pone.0087608.s003\" target=\"_blank\">Table S3</a>. B. Subcellular localization of a fluorescent FlgZ-eCFP fusion protein in the wild-type (F113) strain and in derivatives with high (<i>bifA<sup>-</sup></i>) and low (<i>wspR<sup>-</sup>sadC<sup>-</sup></i>) c-di-GMP levels. The fusion protein is localized at the cellular pole in the <i>bifA</i> mutant, but patch distributed in the <i>wspRsadC</i> double mutant. An intermediate situation is observed in the wild-type strain. Representative confocal micrographies of exponentially growing cells in LB medium are shown.</p>", "links"=>[], "tags"=>["biophysics", "Cell motility", "Flagellar motility", "microbiology", "bacteriology", "Bacterial biofilms", "Bacterial pathogens", "Microbial physiology", "Plant microbiology", "Infectious diseases", "Bacterial diseases", "Pseudomonas infections", "subcellular", "localization", "c-digmp"], "article_id"=>925315, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Francisco Martínez-Granero", "Ana Navazo", "Emma Barahona", "Miguel Redondo-Nieto", "Elena González de Heredia", "Irene Baena", "Irene Martín-Martín", "Rafael Rivilla", "Marta Martín"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087608.g004", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FlgZ_subcellular_localization_is_dependent_on_c_diGMP_levels_/925315", "title"=>"FlgZ subcellular localization is dependent on c-diGMP levels.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-04 03:21:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1375131"], "description"=>"<p>A. Relative swimming motility of <i>flgZ</i> mutants in <i>P. fluorescens</i> F113 and <i>P. putida</i> KT2440 backgrounds. B. Relative swimming motility of <i>P. fluorescens</i> F113 derivatives harboring plasmids which overexpress the F113 (p<i>flgZ</i><sub>F113</sub>) and KT2440 (p<i>flgZ</i><sub>2440</sub>) <i>flgZ</i> genes. C. Relative swimming motility of <i>P. putida</i> KT2440 derivatives harboring plasmids which overexpress the F113 (p<i>flgZ</i><sub>F113</sub>) and KT2440 (p<i>flgZ</i><sub>2440</sub>) <i>flgZ</i> genes. In B and C wild-type strains harbored the empty vector pVLT31 and <i>flgZ</i> expression was induced with 1mM IPTG. Swimming motility was determined by haloes diameter in SA plates with 0.3% agar, 24 hours after inoculation. Averages of three triplicated experiments plus standard deviation are shown. Different letters indicate statistically significant differences (p<0.05). Relative motility of each strain is also shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087608#pone.0087608.s003\" target=\"_blank\">Table S3</a>.</p>", "links"=>[], "tags"=>["biophysics", "Cell motility", "Flagellar motility", "microbiology", "bacteriology", "Bacterial biofilms", "Bacterial pathogens", "Microbial physiology", "Plant microbiology", "Infectious diseases", "Bacterial diseases", "Pseudomonas infections", "implicated"], "article_id"=>925314, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Francisco Martínez-Granero", "Ana Navazo", "Emma Barahona", "Miguel Redondo-Nieto", "Elena González de Heredia", "Irene Baena", "Irene Martín-Martín", "Rafael Rivilla", "Marta Martín"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087608.g003", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_flgZ_is_implicated_in_the_regulation_of_swimming_motility_/925314", "title"=>"<i>flgZ</i> is implicated in the regulation of swimming motility.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-04 03:21:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1375134"], "description"=>"<p>Relative swimming motility of <i>P. fluorescens</i> F113 mutants affected in genes encoding diguanylate cyclases (<i>wspR</i>, <i>sadC</i>), phosphodiesterase (<i>bifA</i>) and putative c-di-GMP sensing and signal transduction proteins (<i>flgZ</i> and <i>sadB</i>). The <i>flgZ</i> mutation did not suppress the swimming phenotypes of <i>wspR</i>, <i>sadC</i> and <i>bifA</i> mutants. Swimming motility was determined by haloes diameter in SA plates with 0.3% agar, 24 hours after inoculation. Averages of three triplicated experiments plus standard deviation are shown. Different letters indicate statistically significant differences (p<0.05). Relative motility of each strain is also shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087608#pone.0087608.s003\" target=\"_blank\">Table S3</a>.</p>", "links"=>[], "tags"=>["biophysics", "Cell motility", "Flagellar motility", "microbiology", "bacteriology", "Bacterial biofilms", "Bacterial pathogens", "Microbial physiology", "Plant microbiology", "Infectious diseases", "Bacterial diseases", "Pseudomonas infections", "modulate", "motility", "independently"], "article_id"=>925317, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Francisco Martínez-Granero", "Ana Navazo", "Emma Barahona", "Miguel Redondo-Nieto", "Elena González de Heredia", "Irene Baena", "Irene Martín-Martín", "Rafael Rivilla", "Marta Martín"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087608.g005", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_c_di_GMP_levels_modulate_swimming_motility_independently_of_FlgZ_/925317", "title"=>"c-di-GMP levels modulate swimming motility independently of FlgZ.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-04 03:21:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1375130"], "description"=>"<p>A. <i>flgZ</i> is co-transcribed with the <i>flgMN</i> genes. RT-PCR analysis of cDNA from total RNA, obtained from <i>P. fluorescens</i> cultures in LB medium at O.D.<sub>600</sub> 0.8. Primers were designed to amplify the indicated fragments. Lanes 1 to 3 correspond to the numbered fragments. Lane 4 is a negative control which contains RNA that was not subjected to retrotranscription. The same expression pattern was found in cells growing in SA medium. B. <i>flgZ</i> expression is regulated by the flagellar regulatory genes <i>fliA</i> and <i>fleQ</i>. qPCR analysis of cDNA from total RNA, obtained from cultures of the wild type F113 strain and mutants affected in the <i>fliA</i> and <i>fleQ</i> genes in LB medium at O.D.<sub>600</sub> 0.8. Primers used amplified an internal region of the <i>flgZ</i> gene. Expression relative to F113 is shown as average plus standard deviation. Different letters indicates statistically significant differences (p<0.05).</p>", "links"=>[], "tags"=>["biophysics", "Cell motility", "Flagellar motility", "microbiology", "bacteriology", "Bacterial biofilms", "Bacterial pathogens", "Microbial physiology", "Plant microbiology", "Infectious diseases", "Bacterial diseases", "Pseudomonas infections", "flagellar"], "article_id"=>925313, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Francisco Martínez-Granero", "Ana Navazo", "Emma Barahona", "Miguel Redondo-Nieto", "Elena González de Heredia", "Irene Baena", "Irene Martín-Martín", "Rafael Rivilla", "Marta Martín"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087608.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_flgZ_is_a_flagellar_gene_/925313", "title"=>"<i>flgZ</i> is a flagellar gene.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-04 03:21:49"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"14", "full-text"=>"16", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"5"}
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  • {"unique-ip"=>"16", "full-text"=>"14", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"20", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"7"}
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  • {"unique-ip"=>"14", "full-text"=>"20", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"10", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"10"}
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  • {"unique-ip"=>"9", "full-text"=>"12", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"15", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2017", "month"=>"2"}
  • {"unique-ip"=>"6", "full-text"=>"3", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"3"}
  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"4"}
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  • {"unique-ip"=>"4", "full-text"=>"6", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2017", "month"=>"7"}
  • {"unique-ip"=>"4", "full-text"=>"4", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"11", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"8"}
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