C-di-GMP Regulates Motile to Sessile Transition by Modulating MshA Pili Biogenesis and Near-Surface Motility Behavior in Vibrio cholerae
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{"title"=>"C-di-GMP Regulates Motile to Sessile Transition by Modulating MshA Pili Biogenesis and Near-Surface Motility Behavior in Vibrio cholerae", "type"=>"journal", "authors"=>[{"first_name"=>"Christopher J.", "last_name"=>"Jones", "scopus_author_id"=>"55474278600"}, {"first_name"=>"Andrew", "last_name"=>"Utada", "scopus_author_id"=>"8928487300"}, {"first_name"=>"Kimberly R.", "last_name"=>"Davis", "scopus_author_id"=>"37661285000"}, {"first_name"=>"Wiriya", "last_name"=>"Thongsomboon", "scopus_author_id"=>"55561887000"}, {"first_name"=>"David", "last_name"=>"Zamorano Sanchez", "scopus_author_id"=>"15754471000"}, {"first_name"=>"Vinita", "last_name"=>"Banakar", "scopus_author_id"=>"54935221200"}, {"first_name"=>"Lynette", "last_name"=>"Cegelski", "scopus_author_id"=>"6506847519"}, {"first_name"=>"Gerard C.L.", "last_name"=>"Wong", "scopus_author_id"=>"7402527046"}, {"first_name"=>"Fitnat H.", "last_name"=>"Yildiz", "scopus_author_id"=>"7006760654"}], "year"=>2015, "source"=>"PLoS Pathogens", "identifiers"=>{"scopus"=>"2-s2.0-84946056908", "sgr"=>"84946056908", "issn"=>"15537374", "isbn"=>"1553-7374 (Electronic) 1553-7366 (Linking)", "pmid"=>"26505896", "doi"=>"10.1371/journal.ppat.1005068", "pui"=>"606740879"}, "id"=>"1ae3cfc2-b688-31cd-92c1-dd0e7d78515e", "abstract"=>"In many bacteria, including Vibrio cholerae, cyclic dimeric guanosine monophosphate (c-di- GMP) controls the motile to biofilm life style switch. Yet, little is known about how this occurs. In this study, we report that changes in c-di-GMP concentration impact the biosynthesis of the MshA pili, resulting in altered motility and biofilm phenotypes in V. cholerae. Previously, we reported that cdgJ encodes a c-di-GMP phosphodiesterase and a ΔcdgJ mutant has reduced motility and enhanced biofilm formation. Here we show that loss of the genes required for the mannose-sensitive hemagglutinin (MshA) pilus biogenesis restores motility in the ΔcdgJ mutant. Mutations of the predicted ATPase proteins mshE or pilT, responsible for polymerizing and depolymerizing MshA pili, impair near surface motility behavior and initial surface attachment dynamics. A ΔcdgJ mutant has enhanced surface attachment, while the ΔcdgJmshA mutant phenocopies the high motility and low attachment phenotypes observed in a ΔmshA strain. Elevated concentrations of c-di-GMP enhance surface MshA pilus production. MshE, but not PilT binds c-di-GMP directly, establishing a mechanism for c-di-GMP signaling input in MshA pilus production. Collectively, our results suggest that the dynamic nature of the MshA pilus established by the assembly and disassembly of pilin subunits is essential for transition from the motile to sessile lifestyle and that c-di-GMP affects MshA pilus assembly and function through direct interactions with the MshE ATPase.", "link"=>"http://www.mendeley.com/research/cdigmp-regulates-motile-sessile-transition-modulating-msha-pili-biogenesis-nearsurface-motility-beha", "reader_count"=>62, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>3, "Researcher"=>12, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>20, "Student > Postgraduate"=>2, "Student > Master"=>12, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>3, "Researcher"=>12, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>20, "Student > Postgraduate"=>2, "Student > Master"=>12, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>13, "Agricultural and Biological Sciences"=>35, "Medicine and Dentistry"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemical Engineering"=>1, "Chemistry"=>2, "Psychology"=>1, "Immunology and Microbiology"=>5}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>2}, "Psychology"=>{"Psychology"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>35}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>13}, "Unspecified"=>{"Unspecified"=>3}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Chemical Engineering"=>{"Chemical Engineering"=>1}}, "reader_count_by_country"=>{"United States"=>2, "Chile"=>1}, "group_count"=>2}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/2382149"], "description"=>"<p>A, C. The number of surface-adhered, stationary cells were counted at the indicated times after inoculation in a flow cell. B, D. Residence times were measured for each cell that interacted with the surface during three high-speed movies of 82s in length within the first 15 minutes post inoculation.</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586784, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.g004", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Early_surface_interactions_are_reduced_in_mshA_mshE_and_pilT_mutants_and_increased_in_cdgJ_mutant_/1586784", "title"=>"Early surface interactions are reduced in <i>mshA</i>, <i>mshE</i>, and <i>pilT</i> mutants and increased in <i>cdgJ</i> mutant.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382170"], "description"=>"<p>A. Isothermal Calorimetry was performed on purified proteins to determine the binding affinity for c-di-GMP. Three independent runs were included in the analysis, one representative run is depicted. VpsT is included as a positive control for c-di-GMP binding. B. Fluorescence thermal shift was utilized to investigate domains of MshE responsible for binding c-di-GMP. Fluorescence melt curves are displayed in the presence of 2mM ATP or c-di-GMP. Three independent experiments were performed in triplicate, with one representative experiment shown. C. Midpoints of melt curves are displayed in the presence of 2mM nucleotides or a buffer control. Three independent experiments were performed in triplicate, Significance was determined with a Oneway ANOVA followed by Dunnett’s Multiple Comparison test comparing to Buffer control. (*** p≤0.001, all others not significant).</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586797, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.g008", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MshE_binds_c_di_GMP_/1586797", "title"=>"MshE binds c-di-GMP.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382143"], "description"=>"<p>The operon encoding genes required for assembly and secretion (<i>mshI-F)</i> is displayed in yellow and the operon encoding pilus structural components <i>mshB-Q</i> is displayed in blue. Transposon insertions are indicated by red arrowheads above the operon. There were no identical insertions observed; arrows are shifted up for clarity indicating two insertions near each other.</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586778, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.g001", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_representation_of_the_transposon_insertions_in_the_Msh_operons_/1586778", "title"=>"Schematic representation of the transposon insertions in the Msh operons.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382156"], "description"=>"<p>A. Surface MshA pilin production determined via ELISA with MshA-specific antibody. Two biological replicates were assayed in quadruplicate and normalized to the average of the WT strain. Oneway ANOVA followed by Bonferroni’s Multiple Comparison test. (ns = not significant, * p≤0.05, ** p≤0.01) B. Western blots detect MshA production from cell pellets using α-MshA antibody. Blots were performed in triplicate, with one representative image included. C. Densitometry of Western blots quantified MshA production in various strains. Densitometry analysis of blots was performed in triplicate and detection was normalized to the WT sample on each blot. Statistical significance determined using a one sample t-test to compare to WT. (*** p≤0.001).</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586790, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.g006", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Production_of_surface_MshA_pili_is_MshE_and_PilT_dependent_and_repressed_by_CdgJ_/1586790", "title"=>"Production of surface MshA pili is MshE and PilT dependent and repressed by CdgJ.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382153"], "description"=>"<p>Measured cell tracks at the indicated times post-inoculation. Bacterial populations were imaged for 82s with a 5 ms resolution, and a representative subset of individual cell tracks are presented for clarity. The field of view is 160μm x 160μm. Each line represents the motion of one cell and different colors are used to denote different cells. Non-motile cells have been filtered out of the data set at each time point for clarity.</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586787, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.g005", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mutations_in_mshA_mshE_pilT_and_cdgJ_affect_near_surface_motility_/1586787", "title"=>"Mutations in <i>mshA</i>, <i>mshE</i>, <i>pilT</i>, and <i>cdgJ</i> affect near surface motility.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382178", "https://ndownloader.figshare.com/files/2382179", "https://ndownloader.figshare.com/files/2382180", "https://ndownloader.figshare.com/files/2382181", "https://ndownloader.figshare.com/files/2382182", "https://ndownloader.figshare.com/files/2382183", "https://ndownloader.figshare.com/files/2382184", "https://ndownloader.figshare.com/files/2382185", "https://ndownloader.figshare.com/files/2382186"], "description"=>"<div><p>In many bacteria, including <i>Vibrio cholerae</i>, cyclic dimeric guanosine monophosphate (c-di-GMP) controls the motile to biofilm life style switch. Yet, little is known about how this occurs. In this study, we report that changes in c-di-GMP concentration impact the biosynthesis of the MshA pili, resulting in altered motility and biofilm phenotypes in <i>V</i>. <i>cholerae</i>. Previously, we reported that <i>cdgJ</i> encodes a c-di-GMP phosphodiesterase and a Δ<i>cdgJ</i> mutant has reduced motility and enhanced biofilm formation. Here we show that loss of the genes required for the mannose<i>-</i>sensitive hemagglutinin <i>(</i>MshA<i>)</i> pilus biogenesis restores motility in the Δ<i>cdgJ</i> mutant. Mutations of the predicted ATPase proteins <i>mshE</i> or <i>pilT</i>, responsible for polymerizing and depolymerizing MshA pili, impair near surface motility behavior and initial surface attachment dynamics. A Δ<i>cdgJ</i> mutant has enhanced surface attachment, while the Δ<i>cdgJmshA</i> mutant phenocopies the high motility and low attachment phenotypes observed in a Δ<i>mshA</i> strain. Elevated concentrations of c-di-GMP enhance surface MshA pilus production. MshE, but not PilT binds c-di-GMP directly, establishing a mechanism for c-di-GMP signaling input in MshA pilus production. Collectively, our results suggest that the dynamic nature of the MshA pilus established by the assembly and disassembly of pilin subunits is essential for transition from the motile to sessile lifestyle and that c-di-GMP affects MshA pilus assembly and function through direct interactions with the MshE ATPase.</p></div>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586804, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>["https://dx.doi.org/10.1371/journal.ppat.1005068.s001", "https://dx.doi.org/10.1371/journal.ppat.1005068.s002", "https://dx.doi.org/10.1371/journal.ppat.1005068.s003", "https://dx.doi.org/10.1371/journal.ppat.1005068.s004", "https://dx.doi.org/10.1371/journal.ppat.1005068.s005", "https://dx.doi.org/10.1371/journal.ppat.1005068.s006", "https://dx.doi.org/10.1371/journal.ppat.1005068.s007", "https://dx.doi.org/10.1371/journal.ppat.1005068.s008", "https://dx.doi.org/10.1371/journal.ppat.1005068.s009"], "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/C_di_GMP_Regulates_Motile_to_Sessile_Transition_by_Modulating_MshA_Pili_Biogenesis_and_Near_Surface_Motility_Behavior_in_Vibrio_cholerae_/1586804", "title"=>"C-di-GMP Regulates Motile to Sessile Transition by Modulating MshA Pili Biogenesis and Near-Surface Motility Behavior in <i>Vibrio cholerae</i>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382173"], "description"=>"<p>Extragenic suppressors of Δ<i>cdgJ</i> motility phenotype.</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586800, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.t001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Extragenic_suppressors_of_cdgJ_motility_phenotype_/1586800", "title"=>"Extragenic suppressors of Δ<i>cdgJ</i> motility phenotype.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382146"], "description"=>"<p>A. Three-dimensional biofilm structures of WT, Δ<i>mshA</i>, Δ<i>mshE</i>, Δ<i>pilT</i>, and Δ<i>pilU</i> strains formed 24 h post inoculation in a flow cell system. Scale bar represents 20μm. B. COMSTAT quantitative analysis of biomass, average thickness, and surface coverage of biofilms from A. Three images from each of two independent experiments were analyzed. Significance was determined with a Oneway ANOVA followed by Dunnett’s Multiple Comparison test comparing to WT. (ns = not significant, *** p≤0.001) C. Three-dimensional biofilm structures of WT, Δ<i>cdgJ</i>, Δ<i>cdgJmshA</i>, Δ<i>cdgJmshE</i>, and Δ<i>cdgJpilT</i>, strains formed after 24 h post inoculation in a flow cell system. Scale bar represents 20μm. D. COMSTAT quantitative analysis of biomass, average thickness, and surface coverage of biofilms from C. Three images from each of two independent experiments were analyzed. Significance was determined with a Oneway ANOVA followed by Dunnett’s Multiple Comparison test comparing to Δ<i>cdgJ</i>. (ns = not significant, * p≤0.05, *** p≤0.001).</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586781, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.g003", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_mshA_mshE_and_pilT_mutants_are_impaired_in_biofilm_formation_/1586781", "title"=>"<i>mshA</i>, <i>mshE</i>, and <i>pilT</i> mutants are impaired in biofilm formation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382171"], "description"=>"<p>Expression of the DGC VCA0956 was induced with varying amounts of IPTG. Intracellular c-di-GMP level was determined with LC-MS/MS and normalized to total protein in the extract (grey bars, right Y axis). Four independent samples were used for each induction condition. Oneway ANOVA followed by Dunnett’s Multiple Comparison Test determined significance. Surface MshA pilin was determined by ELISA (black bars, left Y axis). Three biological replicates were tested in triplicate. Results were normalized to MshA production in the WT strain. Surface MshA pili were significantly increased in all conditions compared to WT (Oneway ANOVA, Dunnett’s Multiple Comparison Test.) Also, there was a significant correlation between increased c-di-GMP and increased surface MshA pili (Pearson correlation, p = 0.0025, R<sup>2</sup> = 0.9199).</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586798, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.g009", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MshA_pilus_production_correlates_with_c_di_GMP_/1586798", "title"=>"MshA pilus production correlates with c-di-GMP.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382168"], "description"=>"<p>Transmission Electron Microscopy of negative stained cells in exponential growth reveals MshA pili. Scale bars included for reference. Representative image presented of at least 8 images analyzed for each strain. Right panel is increased magnification to clearly visualize pili.</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586795, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.g007", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MshA_pilus_production_is_altered_in_an_mshE_mutant_/1586795", "title"=>"MshA pilus production is altered in an <i>mshE</i> mutant.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-10-27 04:11:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/2382145"], "description"=>"<p>A. The diameters of migration zones of the WT and mutants were measured after 16 h of incubation at 30°C on LB soft agar motility plates and normalized to the motility of the WT strain. Four biological replicates were performed in quadruplicate. Statistical significance determined with Oneway ANOVA followed by Dunnett’s multiple comparison test comparing to the WT strain. (ns—not significant, *** p≤0.001) B. Expression of <i>mshA</i> reduces motility in soft agar. The diameters of migration zones were measured after 16 hours of growth at 30°C with or without IPTG induction and normalized to the motility of the WT strain. Two biological replicates were performed in triplicate. Induced strains compared to uninduced strains with Student’s t-test. (ns—not significant, *** p≤0.001).</p>", "links"=>[], "tags"=>["Vibrio cholerae", "Modulating MshA Pili Biogenesis", "surface attachment dynamics", "ATPase proteins mshE", "surface MshA pilus production", "MshA pilus assembly", "MshA pilus production", "cyclic dimeric guanosine monophosphate", "Δ mshA strain", "surface motility behavior", "biofilm life style switch", "Δ cdgJ", "depolymerizing MshA pili"], "article_id"=>1586780, "categories"=>["Biological Sciences"], "users"=>["Christopher J. Jones", "Andrew Utada", "Kimberly R. Davis", "Wiriya Thongsomboon", "David Zamorano Sanchez", "Vinita Banakar", "Lynette Cegelski", "Gerard C. L. Wong", "Fitnat H. Yildiz"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1005068.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mutations_in_mshA_mshE_and_pilT_increase_motility_/1586780", "title"=>"Mutations in <i>mshA</i>, <i>mshE</i>, and <i>pilT</i> increase motility.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-10-27 04:11:33"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"12", "full-text"=>"15", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2018", "month"=>"5"}
  • {"unique-ip"=>"17", "full-text"=>"18", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"5", "cited-by"=>"0", "year"=>"2018", "month"=>"7"}
  • {"unique-ip"=>"11", "full-text"=>"27", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"6"}
  • {"unique-ip"=>"12", "full-text"=>"22", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
  • {"unique-ip"=>"10", "full-text"=>"12", "pdf"=>"13", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"8"}
  • {"unique-ip"=>"13", "full-text"=>"15", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
  • {"unique-ip"=>"23", "full-text"=>"29", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"11", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
  • {"unique-ip"=>"17", "full-text"=>"33", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
  • {"unique-ip"=>"22", "full-text"=>"38", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"17", "full-text"=>"25", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"20", "full-text"=>"154", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"34", "full-text"=>"47", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"10", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"9", "full-text"=>"11", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"8", "full-text"=>"8", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"12", "full-text"=>"11", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"9", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}
  • {"unique-ip"=>"22", "full-text"=>"20", "pdf"=>"9", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"12"}
  • {"unique-ip"=>"33", "full-text"=>"20", "pdf"=>"26", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2020", "month"=>"2"}
  • {"unique-ip"=>"20", "full-text"=>"24", "pdf"=>"10", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"3"}
  • {"unique-ip"=>"11", "full-text"=>"12", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"4"}
  • {"unique-ip"=>"14", "full-text"=>"20", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"5"}
  • {"unique-ip"=>"17", "full-text"=>"20", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"6"}
  • {"unique-ip"=>"5", "full-text"=>"3", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"7"}
  • {"unique-ip"=>"20", "full-text"=>"21", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"10", "cited-by"=>"0", "year"=>"2020", "month"=>"8"}
  • {"unique-ip"=>"5", "full-text"=>"7", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"9"}
  • {"unique-ip"=>"20", "full-text"=>"17", "pdf"=>"9", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2020", "month"=>"10"}

Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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