The NDR/LATS Family Kinase Cbk1 Directly Controls Transcriptional Asymmetry
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{"title"=>"The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry", "type"=>"journal", "authors"=>[{"first_name"=>"Emily", "last_name"=>"Mazanka", "scopus_author_id"=>"24724943200"}, {"first_name"=>"Jess", "last_name"=>"Alexander", "scopus_author_id"=>"56666520200"}, {"first_name"=>"Brian J.", "last_name"=>"Yeh", "scopus_author_id"=>"7005394391"}, {"first_name"=>"Patrick", "last_name"=>"Charoenpong", "scopus_author_id"=>"24723670600"}, {"first_name"=>"Drew M.", "last_name"=>"Lowery", "scopus_author_id"=>"35555606100"}, {"first_name"=>"Michael", "last_name"=>"Yaffe", "scopus_author_id"=>"7101980618"}, {"first_name"=>"Eric L.", "last_name"=>"Weiss", "scopus_author_id"=>"7402352569"}], "year"=>2008, "source"=>"PLoS Biology", "identifiers"=>{"doi"=>"10.1371/journal.pbio.0060203", "pui"=>"352236835", "issn"=>"15449173", "scopus"=>"2-s2.0-50249177622", "sgr"=>"50249177622", "pmid"=>"18715118", "isbn"=>"1545-7885 (Electronic)"}, "id"=>"bb61b7cd-31cd-3984-88c3-fab4d4fa40f7", "abstract"=>"Cell fate can be determined by asymmetric segregation of gene expression regulators. In the budding yeast Saccharomyces cerevisiae, the transcription factor Ace2 accumulates specifically in the daughter cell nucleus, where it drives transcription of genes that are not expressed in the mother cell. The NDR/LATS family protein kinase Cbk1 is required for Ace2 segregation and function. Using peptide scanning arrays, we determined Cbk1's phosphorylation consensus motif, the first such unbiased approach for an enzyme of this family, showing that it is a basophilic kinase with an unusual preference for histidine -5 to the phosphorylation site. We found that Cbk1 phosphorylates such sites in Ace2, and that these modifications are critical for Ace2's partitioning and function. Using proteins marked with GFP variants, we found that Ace2 moves from isotropic distribution to the daughter cell nuclear localization, well before cytokinesis, and that the nucleus must enter the daughter cell for Ace2 accumulation to occur. We found that Cbk1, unlike Ace2, is restricted to the daughter cell. Using both in vivo and in vitro assays, we found that two critical Cbk1 phosphorylations block Ace2's interaction with nuclear export machinery, while a third distal modification most likely acts to increase the transcription factor's activity. Our findings show that Cbk1 directly controls Ace2, regulating the transcription factor's activity and interaction with nuclear export machinery through three phosphorylation sites. Furthermore, Cbk1 exhibits a novel specificity that is likely conserved among related kinases from yeast to metazoans. Cbk1 is functionally restricted to the daughter cell, and cannot diffuse from the daughter to the mother. In addition to providing a mechanism for Ace2 segregation, these findings show that an isotropically distributed cell fate determinant can be asymmetrically partitioned in cytoplasmically contiguous cells through spatial segregation of a regulating protein kinase.", "link"=>"http://www.mendeley.com/research/ndrlats-family-kinase-cbk1-directly-controls-transcriptional-asymmetry", "reader_count"=>60, "reader_count_by_academic_status"=>{"Researcher"=>24, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>15, "Student > Postgraduate"=>1, "Student > Master"=>5, "Student > Bachelor"=>8, "Professor"=>5}, "reader_count_by_user_role"=>{"Researcher"=>24, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>15, "Student > Postgraduate"=>1, "Student > Master"=>5, "Student > Bachelor"=>8, "Professor"=>5}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>11, "Agricultural and Biological Sciences"=>42, "Medicine and Dentistry"=>4, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Physics and Astronomy"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>42}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>11}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United States"=>1, "Brazil"=>1, "France"=>1, "Germany"=>1, "Spain"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/924328"], "description"=>"<div><p>(A) Quantification of cell separation defect in <i>ace2</i> mutant strains, shown as the percentage of groups of cells in the population in clumps of the indicated number of connected cells. The number of connected cells per group was counted in three separate trials (<i>n</i> = approximately 200 groups per trial) for wild-type (WT), <i>ace2-2A</i>, <i>ace2-3A</i>, and <i>ace2Δ</i> strains; standard deviation of the mean fraction of the population per group for the three trials is shown (analysis of other alleles, <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0060203#pbio-0060203-sg003\" target=\"_blank\">Figure S3</a>).</p>\n <p>(B) Localization of wild-type Ace2-GFP, double-mutant <i>ace2-2A-GFP</i>, triple-mutant <i>ace2-3A</i>-GFP, and wild-type Ace2-GFP in a <i>cbk1Δ</i> strain. Mother cells were labeled with rhodamine-ConA (red); daughter cells were unlabeled. Blue brackets indicate mother–daughter pairs. Cells carrying <i>ace2-3A</i>-GFP accumulate as large clusters, and <i>ace2-3A</i>-GFP localizes to both mother and daughter nuclei. Images were all contrast enhanced to the same setting for direct comparison. Scale bar indicates 5 μm.</p>\n <p>(C) Quantification of GFP fluorescence intensity in 40 individual nuclei of large budded mother–daughter paired cells carrying <i>ace2-3A</i>-GFP, compared with <i>ace2-2A-GFP</i> and Ace2-GFP in both wild-type and <i>cbk1Δ</i> strains. Error bars indicate standard error of the mean.</p>\n <p>(D) Expression of Ace2 target genes <i>CTS1</i> and <i>DSE1</i> measured by quantitative RT-PCR demonstrated that <i>ace2-3A</i> was defective for transcriptional activity. Shown are the average values and standard deviation of three independent trials. Gene expression in all <i>ace2</i> mutant strains is shown in <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0060203#pbio-0060203-sg004\" target=\"_blank\">Figure S4</a>.</p>\n <p>(E) Analysis of DNA binding to <i>DSE1</i> promoter shows that <i>ace2-3A</i>-HA fails to bind DNA targets, comparable to the loss of binding seen in an Ace2-HA <i>cbk1Δ</i> strain. Standard error of the mean of at least six replicates from three independent experiments is shown. Solid line indicates wild-type level of binding; dashed line denotes untagged control (as shown in <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0060203#pbio-0060203-g002\" target=\"_blank\">Figure 2</a>).</p></div>", "links"=>[], "tags"=>["amino", "acids", "s436", "ace2", "localization"], "article_id"=>594772, "categories"=>["Biochemistry", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Emily Mazanka", "Jess Alexander", "Brian J Yeh", "Patrick Charoenpong", "Drew M Lowery", "Michael Yaffe", "Eric L Weiss"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0060203.g005", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phosphoacceptor_Amino_Acids_S122_S137_and_S436_Are_Required_for_Proper_Ace2_Localization_and_Function_In_Vivo_/594772", "title"=>"Phosphoacceptor Amino Acids S122, S137, and S436 Are Required for Proper Ace2 Localization and Function In Vivo", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-08-19 01:19:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/924438"], "description"=>"<div><p>(A) A biotinylated peptide encompassing residues 122–150 of Ace2 was sufficient to interact with Crm1 in an in vitro pulldown experiment. Peptides containing phosphoserine at positions 122 and 137 bound Crm1 more weakly; however, wild-type (WT) binding was restored by pretreatment of the peptides with λ-phosphatase. A F127V mutation in Ace2 also weakened the interaction with Crm1.</p>\n <p>(B) Quantification of cell separation in <i>ace2-F127V-3A</i> and <i>ace2-S122D/S137D</i> strains compared to <i>ace2-3A</i> and <i>ace2-F127V cbk1Δ</i>. <i>cbk1Δ</i> quantification is shown in <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0060203#pbio-0060203-sg003\" target=\"_blank\">Figure S3</a>. Three independent trials of 200 groups of cells were counted and binned into different clump sizes as shown. Percentage of the population found in each clump size and standard deviations are shown. Addition of the F127V mutation suppresses the cell separation defect of <i>ace2-3A</i> cells.</p>\n <p>(C) Fluorescence intensity of each GFP-tagged Ace2 allele was quantified in approximately 40 nuclei of large budded mother–daughter pairs. <i>ace2-F127V-3A</i>-GFP shows increased nuclear accumulation compared to <i>ace2-3A</i>-GFP. Standard error of the mean is shown.</p>\n <p>(D) Quantitative RT-PCR of Ace2 target genes <i>CTS1</i> and <i>DSE1</i> is shown for each Ace2 allele. Mutation of F127V in the <i>ace2-3A</i> background increases transcriptional activity. Shown is the average of three trials with error bars denoting standard deviation.</p></div>", "links"=>[], "tags"=>["nes", "suppresses"], "article_id"=>594887, "categories"=>["Biochemistry", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Emily Mazanka", "Jess Alexander", "Brian J Yeh", "Patrick Charoenpong", "Drew M Lowery", "Michael Yaffe", "Eric L Weiss"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0060203.g006", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mutation_of_ace2_3A_NES_Suppresses_Loss_of_Function_Phenotype_/594887", "title"=>"Mutation of <i>ace2-3A</i> NES Suppresses Loss of Function Phenotype", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-08-19 01:21:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/924136"], "description"=>"<p>Mother/daughter asymmetry of Ace2-GFP (A) and Cbk1-GFP (B) following block of nuclear export. Left panels: representative cells ± 30 min treatment with leptomycin B (LMB). Scale bars indicate 5 μm. Graphs: percentage of cells exhibiting mother-only, daughter-only, or symmetric mother/daughter nuclear localization ± LMB, <i>n</i> ≥ 50 nuclei per trial.</p>", "links"=>[], "tags"=>["partitioned"], "article_id"=>594584, "categories"=>["Biochemistry", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Emily Mazanka", "Jess Alexander", "Brian J Yeh", "Patrick Charoenpong", "Drew M Lowery", "Michael Yaffe", "Eric L Weiss"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0060203.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cbk1_Is_Partitioned_to_the_Daughter_Cell_/594584", "title"=>"Cbk1 Is Partitioned to the Daughter Cell", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-08-19 01:16:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/455698", "https://ndownloader.figshare.com/files/455740", "https://ndownloader.figshare.com/files/455778", "https://ndownloader.figshare.com/files/455822", "https://ndownloader.figshare.com/files/455865"], "description"=>"<div><p>Cell fate can be determined by asymmetric segregation of gene expression regulators. In the budding yeast Saccharomyces cerevisiae, the transcription factor Ace2 accumulates specifically in the daughter cell nucleus, where it drives transcription of genes that are not expressed in the mother cell. The NDR/LATS family protein kinase Cbk1 is required for Ace2 segregation and function. Using peptide scanning arrays, we determined Cbk1′s phosphorylation consensus motif, the first such unbiased approach for an enzyme of this family, showing that it is a basophilic kinase with an unusual preference for histidine −5 to the phosphorylation site. We found that Cbk1 phosphorylates such sites in Ace2, and that these modifications are critical for Ace2′s partitioning and function. Using proteins marked with GFP variants, we found that Ace2 moves from isotropic distribution to the daughter cell nuclear localization, well before cytokinesis, and that the nucleus must enter the daughter cell for Ace2 accumulation to occur. We found that Cbk1, unlike Ace2, is restricted to the daughter cell. Using both in vivo and in vitro assays, we found that two critical Cbk1 phosphorylations block Ace2′s interaction with nuclear export machinery, while a third distal modification most likely acts to increase the transcription factor's activity. Our findings show that Cbk1 directly controls Ace2, regulating the transcription factor's activity and interaction with nuclear export machinery through three phosphorylation sites. Furthermore, Cbk1 exhibits a novel specificity that is likely conserved among related kinases from yeast to metazoans. Cbk1 is functionally restricted to the daughter cell, and cannot diffuse from the daughter to the mother. In addition to providing a mechanism for Ace2 segregation, these findings show that an isotropically distributed cell fate determinant can be asymmetrically partitioned in cytoplasmically contiguous cells through spatial segregation of a regulating protein kinase.</p> </div>", "links"=>[], "tags"=>["kinase", "cbk1", "controls", "transcriptional", "asymmetry"], "article_id"=>149754, "categories"=>["Biochemistry", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Emily Mazanka", "Jess Alexander", "Brian J Yeh", "Patrick Charoenpong", "Drew M Lowery", "Michael Yaffe", "Eric L Weiss"], "doi"=>["https://dx.doi.org/10.1371/journal.pbio.0060203.sg001", "https://dx.doi.org/10.1371/journal.pbio.0060203.sg002", "https://dx.doi.org/10.1371/journal.pbio.0060203.sg003", "https://dx.doi.org/10.1371/journal.pbio.0060203.sg004", "https://dx.doi.org/10.1371/journal.pbio.0060203.sg005"], "stats"=>{"downloads"=>24, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/The_NDR_LATS_Family_Kinase_Cbk1_Directly_Controls_Transcriptional_Asymmetry_/149754", "title"=>"The NDR/LATS Family Kinase Cbk1 Directly Controls Transcriptional Asymmetry", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2008-08-19 02:42:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/924237"], "description"=>"<div><p>(A) Cbk1 kinase domain purified from E. coli was incubated with the indicated array of biotinylated peptide libraries in the presence of γ-<sup>32</sup>P-ATP, transferred to a strepavidin membrane, and kinase activity was analyzed by autoradiography. Numbers at the top of each column indicate positions relative to the phosphoacceptor S/T residue; rows contain specific amino acids in those positions. The optimal Cbk1 consensus sequence is HXRRX(S/T) as determined by quantification of the most highly phosphorylated peptide pools in the array.</p>\n <p>(B) Schematic diagram of Ace2 protein showing location of Cbk1 phosphorylation consensus sequences.</p>\n <p>(C) In vitro phosphorylation of N-terminally GST-tagged Ace2 fragments by immunoprecipitated Cbk1-HA. Combined Ala substitution of residues S113, S122, and S137 abolished <sup>32</sup>P labeling of the Ace2 fragment (lane 4). His at position 117 is required for phosphorylation of S122 (lane 5). Protein levels for GST-Ace2 and Cbk1-HA were confirmed to be similar for all reactions by immunoblotting against GST and HA. WT, wild-type.</p></div>", "links"=>[], "tags"=>["exhibits", "phosphorylation", "motif"], "article_id"=>594683, "categories"=>["Biochemistry", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Emily Mazanka", "Jess Alexander", "Brian J Yeh", "Patrick Charoenpong", "Drew M Lowery", "Michael Yaffe", "Eric L Weiss"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0060203.g004", "stats"=>{"downloads"=>1, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cbk1_Exhibits_Phosphorylation_Site_Preference_for_a_Basic_Motif_with_His_at_the_8722_5_Position_/594683", "title"=>"Cbk1 Exhibits Phosphorylation Site Preference for a Basic Motif with His at the −5 Position", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-08-19 01:18:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/923920"], "description"=>"<div><p>(A) Time-lapse microscopy of live cells expressing Ace2-GFP and Myo1-Cherry. Ace2-GFP localization to daughter nuclei is visible prior to disappearance of Myo1-Cherry from the bud neck.</p>\n <p>(B) Nuclear accumulation of Ace2-GFP in <i>arp1Δ bub2Δ</i> cells that fail to segregate the nucleus to the daughter. Ace2-GFP shown in left panel, and nuclear DNA stained with Hoechst shown in right panel. Ace2-GFP localizes normally in cells in which the daughter cell receives a nucleus (cell a, left panel). No nuclear accumulation of Ace2-GFP is observed in cells in which the nucleus divides entirely in the mother cell (cells b and c, left panel). Scale bar indicates 5 μm.</p>\n <p>(C) Quantification of cells shown in (B). Ace2-GFP nuclear accumulation in budded cells with two nuclei in mother cells (left graph) was scored as localized to one of the two nuclei (1), both nuclei (2), or unlocalized, <i>n</i> = 43 cells. For comparison, Ace2-GFP localization in budded cells with properly segregated nuclei (right graph) was scored as daughter nucleus only (D), mother nucleus only (M), both nuclei (M+D), or unlocalized, <i>n</i> = 53 cells.</p></div>", "links"=>[], "tags"=>["localization", "nuclei", "occurs", "cytokinesis", "requires", "nucleus"], "article_id"=>594362, "categories"=>["Biochemistry", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Emily Mazanka", "Jess Alexander", "Brian J Yeh", "Patrick Charoenpong", "Drew M Lowery", "Michael Yaffe", "Eric L Weiss"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0060203.g001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ace2_Localization_to_Daughter_Nuclei_Occurs_Prior_to_Cytokinesis_and_Requires_Movement_of_the_Nucleus_into_the_Daughter_Cell_/594362", "title"=>"Ace2 Localization to Daughter Nuclei Occurs Prior to Cytokinesis and Requires Movement of the Nucleus into the Daughter Cell", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-08-19 01:12:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/924048"], "description"=>"<div><p>(A) Amino acid composition of Ace2 zinc finger domains are shown. Red letters denote residues predicted to participate in DNA binding. Ala substitutions were made at all eight of these sites for microscopy, ChIP, and quantitative RT-PCR analysis.</p>\n <p>(B) ChIP analysis of Ace2 binding to <i>DSE1</i> promoter shows that <i>ace2-8Z</i>-HA cannot bind its DNA target. Standard error of the mean of at least six replicates from three independent experiments is shown. WT, wild-type.</p>\n <p>(C) Quantitative RT-PCR measurement of <i>CTS1</i> and <i>DSE1</i> transcription indicates that <i>ace2-8Z</i>-GFP cannot drive expression of Ace2 target genes.</p>\n <p>(D) DNA binding-deficient mutant <i>ace2-8Z</i>-GFP is unable to promote cell separation, but localizes to daughter nuclei. Asynchronous cells were pulse labeled with rhodamine-ConA for 10 min to label the cell wall. The cells were then grown for 70 min following the removal of fluorescent lectin. Mother cells are stained in red, daughter cells are unstained. Blue brackets indicate mother–daughter pairs. Scale bars indicate 5 μm.</p></div>", "links"=>[], "tags"=>["daughter-specific", "localization", "dna"], "article_id"=>594495, "categories"=>["Biochemistry", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Emily Mazanka", "Jess Alexander", "Brian J Yeh", "Patrick Charoenpong", "Drew M Lowery", "Michael Yaffe", "Eric L Weiss"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0060203.g002", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ace2_Daughter_Specific_Localization_Does_Not_Require_DNA_Binding_/594495", "title"=>"Ace2 Daughter-Specific Localization Does Not Require DNA Binding", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-08-19 01:14:55"}

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  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"3"}
  • {"unique-ip"=>"5", "full-text"=>"7", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2015", "month"=>"4"}
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  • {"unique-ip"=>"9", "full-text"=>"7", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"1", "cited-by"=>"3", "year"=>"2015", "month"=>"11"}
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  • {"unique-ip"=>"8", "full-text"=>"4", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"11", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"3"}
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  • {"unique-ip"=>"11", "full-text"=>"12", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"5"}
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  • {"unique-ip"=>"2", "full-text"=>"1", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"11"}
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  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"4"}
  • {"unique-ip"=>"1", "full-text"=>"1", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"6"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2017", "month"=>"7"}
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  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"9"}
  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"10"}

Relative Metric

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