Yes-Associated Protein 65 (YAP) Expands Neural Progenitors and Regulates Pax3 Expression in the Neural Plate Border Zone
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{"title"=>"Yes-associated protein 65 (YAP) expands neural progenitors and regulates pax3 expression in the neural plate border zone", "type"=>"journal", "authors"=>[{"first_name"=>"Stephen T.", "last_name"=>"Gee", "scopus_author_id"=>"42061341700"}, {"first_name"=>"Sharon L.", "last_name"=>"Milgram", "scopus_author_id"=>"7003393526"}, {"first_name"=>"Kenneth L.", "last_name"=>"Kramer", "scopus_author_id"=>"7201849073"}, {"first_name"=>"Frank L.", "last_name"=>"Conlon", "scopus_author_id"=>"6701816280"}, {"first_name"=>"Sally A.", "last_name"=>"Moody", "scopus_author_id"=>"7005632454"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-79958128093", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0020309", "pui"=>"361908358", "sgr"=>"79958128093", "issn"=>"19326203", "pmid"=>"21687713"}, "id"=>"54696e1b-06d5-34ce-a74c-6774a842ae48", "abstract"=>"Yes-associated protein 65 (YAP) contains multiple protein-protein interaction domains and functions as both a transcriptional co-activator and as a scaffolding protein. Mouse embryos lacking YAP did not survive past embryonic day 8.5 and showed signs of defective yolk sac vasculogenesis, chorioallantoic fusion, and anterior-posterior (A-P) axis elongation. Given that the YAP knockout mouse defects might be due in part to nutritional deficiencies, we sought to better characterize a role for YAP during early development using embryos that develop externally. YAP morpholino (MO)-mediated loss-of-function in both frog and fish resulted in incomplete epiboly at gastrulation and impaired axis formation, similar to the mouse phenotype. In frog, germ layer specific genes were expressed, but they were temporally delayed. YAP MO-mediated partial knockdown in frog allowed a shortened axis to form. YAP gain-of-function in Xenopus expanded the progenitor populations in the neural plate (sox2(+)) and neural plate border zone (pax3(+)), while inhibiting the expression of later markers of tissues derived from the neural plate border zone (neural crest, pre-placodal ectoderm, hatching gland), as well as epidermis and somitic muscle. YAP directly regulates pax3 expression via association with TEAD1 (N-TEF) at a highly conserved, previously undescribed, TEAD-binding site within the 5' regulatory region of pax3. Structure/function analyses revealed that the PDZ-binding motif of YAP contributes to the inhibition of epidermal and somitic muscle differentiation, but a complete, intact YAP protein is required for expansion of the neural plate and neural plate border zone progenitor pools. These results provide a thorough analysis of YAP mediated gene expression changes in loss- and gain-of-function experiments. Furthermore, this is the first report to use YAP structure-function analyzes to determine which portion of YAP is involved in specific gene expression changes and the first to show direct in vivo evidence of YAP's role in regulating pax3 neural crest expression.", "link"=>"http://www.mendeley.com/research/yesassociated-protein-65-yap-expands-neural-progenitors-regulates-pax3-expression-neural-plate-borde", "reader_count"=>69, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Librarian"=>1, "Researcher"=>15, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>25, "Student > Postgraduate"=>3, "Student > Master"=>6, "Other"=>2, "Student > Bachelor"=>6, "Lecturer"=>2, "Professor"=>5}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Librarian"=>1, "Researcher"=>15, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>25, "Student > Postgraduate"=>3, "Student > Master"=>6, "Other"=>2, "Student > Bachelor"=>6, "Lecturer"=>2, "Professor"=>5}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>11, "Agricultural and Biological Sciences"=>49, "Medicine and Dentistry"=>4, "Neuroscience"=>1, "Physics and Astronomy"=>1, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Neuroscience"=>{"Neuroscience"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>49}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>11}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"United States"=>3, "Guatemala"=>1, "Chile"=>1, "France"=>1, "Portugal"=>1}, "group_count"=>5}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/767834"], "description"=>"<p>(<b>A</b>) The <i>pax3</i>-expressing neural crest progenitor field (NCP) is darker, longer, and/or wider (bracket) on the <i>xyap</i>-injected side. Dorsal view, stage 15. (<b>B</b>) xYAP MO-mediated knockdown (40 ng) eliminated <i>pax3</i> expression in both neural crest progenitors and hatching gland (HG) precursors. Addition of exogenous <i>xyap</i> (YAP MO + <i>xyap</i>) rescued <i>pax3</i> expression in neural crest progenitors (NCP), but not in hatching gland. Dorsal views, stage 17. (<b>C</b>) <i>xyap</i> mRNA injection into the precursor blastomere of the intermediate mesoderm does not cause <i>pax3</i> expansion on the injected side (left panel, dorsal view). Right panel (posterior view, dorsal is down) shows part of the lineage labeled clone (red) denoting the injected side. (<b>D</b>) <i>tead1</i> mRNA injection (100 pg) expands <i>pax3</i>-expressing neural crest progenitors (NCP) at a moderate frequency. <i>xyap</i> mRNA injection (100 pg) rarely expands this population. In combination (<i>tead1</i>/<i>xyap</i>, 100 pg each), this population is expanded in nearly every embryo. The repression of the <i>pax3</i>-expressing hatching gland progenitors (HG) also was greatest when TEAD/xYAP were co-expressed. Dorsal anterior views at stage 16.</p>", "links"=>[], "tags"=>["expands", "neural", "crest"], "article_id"=>438211, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020309.g006", "stats"=>{"downloads"=>8, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_xYAP_expands_pax3_expressing_neural_crest_progenitors_/438211", "title"=>"xYAP expands <i>pax3</i>-expressing neural crest progenitors.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-08 02:16:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/767425"], "description"=>"<p>(<b>A</b>) Time-lapse videomicroscopy shows that zYAP gain-of-function does not alter the timing of gastrulation movements, as evidenced by the progression of epiboly (asterisks mark the fronts of tissue movement around the yolk). (<b>B</b>) zYAP gain-of-function in <i>Danio rerio</i> embryos resulted in head and eye deformities and shortened, malformed body axes. Examples of two different mRNA doses are shown. (<b>C</b>) Injection of <i>Xenopus</i> (x), mouse (m), or human (h) <i>yap</i> mRNAs into <i>Xenopus</i> embryos all showed phenotypes similar to those in zebrafish.</p>", "links"=>[], "tags"=>["xyap", "gain-of-function", "axis"], "article_id"=>437795, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020309.g003", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_zYAP_and_xYAP_gain_of_function_results_in_similar_body_axis_defects_/437795", "title"=>"zYAP and xYAP gain-of-function results in similar body axis defects.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-08 02:09:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/386001", "https://ndownloader.figshare.com/files/386038", "https://ndownloader.figshare.com/files/386077", "https://ndownloader.figshare.com/files/386090"], "description"=>"<div><p>Yes-associated protein 65 (YAP) contains multiple protein-protein interaction domains and functions as both a transcriptional co-activator and as a scaffolding protein. Mouse embryos lacking YAP did not survive past embryonic day 8.5 and showed signs of defective yolk sac vasculogenesis, chorioallantoic fusion, and anterior-posterior (A-P) axis elongation. Given that the YAP knockout mouse defects might be due in part to nutritional deficiencies, we sought to better characterize a role for YAP during early development using embryos that develop externally. YAP morpholino (MO)-mediated loss-of-function in both frog and fish resulted in incomplete epiboly at gastrulation and impaired axis formation, similar to the mouse phenotype. In frog, germ layer specific genes were expressed, but they were temporally delayed. YAP MO-mediated partial knockdown in frog allowed a shortened axis to form. YAP gain-of-function in <em>Xenopus</em> expanded the progenitor populations in the neural plate (<em>sox2<sup>+</sup></em>) and neural plate border zone (<em>pax3<sup>+</sup></em>), while inhibiting the expression of later markers of tissues derived from the neural plate border zone (neural crest, pre-placodal ectoderm, hatching gland), as well as epidermis and somitic muscle. YAP directly regulates <em>pax3</em> expression via association with TEAD1 (N-TEF) at a highly conserved, previously undescribed, TEAD-binding site within the 5′ regulatory region of <em>pax3</em>. Structure/function analyses revealed that the PDZ-binding motif of YAP contributes to the inhibition of epidermal and somitic muscle differentiation, but a complete, intact YAP protein is required for expansion of the neural plate and neural plate border zone progenitor pools. These results provide a thorough analysis of YAP mediated gene expression changes in loss- and gain-of-function experiments. Furthermore, this is the first report to use YAP structure-function analyzes to determine which portion of YAP is involved in specific gene expression changes and the first to show direct <em>in vivo</em> evidence of YAP's role in regulating <em>pax3</em> neural crest expression.</p> </div>", "links"=>[], "tags"=>["yes-associated", "65", "expands", "neural", "progenitors", "regulates"], "article_id"=>136198, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0020309.s001", "https://dx.doi.org/10.1371/journal.pone.0020309.s002", "https://dx.doi.org/10.1371/journal.pone.0020309.s003", "https://dx.doi.org/10.1371/journal.pone.0020309.s004"], "stats"=>{"downloads"=>3, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Yes_Associated_Protein_65_YAP_Expands_Neural_Progenitors_and_Regulates_Pax3_Expression_in_the_Neural_Plate_Border_Zone/136198", "title"=>"Yes-Associated Protein 65 (YAP) Expands Neural Progenitors and Regulates <em>Pax3</em> Expression in the Neural Plate Border Zone", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-06-08 01:43:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/768077"], "description"=>"<p>(<b>A</b>) Cartoons of the xYAP mutants created to determine which protein-protein interaction domain(s) is important for the <i>in vivo</i> gain-of-function phenotypes described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020309#pone-0020309-g004\" target=\"_blank\">Figures 4</a>–<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020309#pone-0020309-g005\" target=\"_blank\"></a><a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020309#pone-0020309-g006\" target=\"_blank\">6</a>. Deletions or mutations are indicated by color loss: the TEAD-binding site (xYAPΔTBS, purple), the LATS phosphorylation site (cActive xYAP, orange), the two WW domains (xYAPΔWW, red), the N-terminus (xYAP, ΔN-term) containing both the hnRNP U and TEAD-binding sites, and the PDZ-binding motif (xYAPΔC-term, fuchsia) at the C-terminus. (<b>B</b>) The percentage of embryos showing expansion of <i>sox2</i>-expressing neural plate cells or expansion of <i>pax3</i>-expressing neural crest progenitor (NCP) cells after injection of each of the mutant forms of xYAP. Note that cActive xYAP, which prevents YAP from leaving the nucleus, is as effective as wild type YAP. However, all other mutant forms reduce this phenotype. Sample sizes are presented in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020309#pone-0020309-t001\" target=\"_blank\">Table 1</a>. (<b>C</b>) The percentage of embryos showing reduced gene expression after injection of each mutant form of xYAP. Deletion of the WW domains or of the PDZ-binding motif interfered the most with repression of <i>pax3</i><sup>+</sup> hatching gland (HG) progenitors. Loss of neural plate differentiation (<i>p27<sup>xic1</sup></i>) and a PPE marker (<i>sox11</i>) were maintained at high frequencies with each xYAP mutant, indicating that interactions at one or more of the remaining domains are sufficient to downregulate these genes. However, xYAP-mediated loss of somitic muscle (<i>myoD</i>) and epidermal (<i>cyto-keratin</i>) differentiation was specifically reduced by deletion of its PDZ-binding motif.</p>", "links"=>[], "tags"=>["deletion", "mutants", "differential"], "article_id"=>438449, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020309.g008", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_xYAP_deletion_mutants_exhibit_differential_activities_/438449", "title"=>"xYAP deletion mutants exhibit differential activities.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-08 02:20:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/767284"], "description"=>"<p>(<b>A</b>) qPCR analysis of mRNA from uninjected, control MO-injected, and xYAP MO-injected <i>Xenopus</i> embryos collected when controls reached stage 10.5/11. <i>brachyury</i>, <i>goosecoid</i>, <i>wnt8</i>, <i>sox11</i>, and <i>sox17</i> mRNA levels were reduced, <i>nodal-related 3</i> (<i>nr3</i>) mRNA levels were increased and <i>siamois</i> mRNA levels remained unchanged in xYAP morphant embryos. (<b>B</b>) <i>In situ</i> characterization of mesoderm gene expression in uninjected, control MO-, and xYAP MO-injected <i>Xenopus</i> embryos. xYAP morphant embryos express each gene in the correct location, but the spatial pattern resembles an earlier developmental stage. For example, <i>brachyury</i> expression in the stage 11 YAP MO embryos is only faintly detected and <i>brachyury</i> expression in the stage 13 YAP MO embryo is indistinguishable from the control stage 11 pattern. <i>chordin</i> expression in the stage 13 YAP MO embryo remains confined to the dorsal blastopore lip (arrow), as is normal at stage 11; it has not elongated with the axial mesoderm as is normal at stage 13. <i>eomesodermin</i> expression in the stage 11 YAP MO embryo remains on the surface in the uninvoluted mesoderm (arrow), whereas in controls, <i>eomesodermin</i>-expressing cells have migrated internally <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020309#pone.0020309-Ryan1\" target=\"_blank\">[21]</a>. In the stage 11 panel, all views are vegetal; in the stage 13 panel, the views of <i>brachyury</i> and <i>vent2</i> embryos and of the YAP MO <i>chordin</i> embryos are vegetal and the remainder are dorsal.</p>", "links"=>[], "tags"=>["markers", "yap", "morphant", "temporally"], "article_id"=>437651, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020309.g002", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Germ_layer_markers_are_expressed_in_YAP_morphant_Xenopus_embryos_but_are_temporally_delayed_/437651", "title"=>"Germ layer markers are expressed in YAP morphant <i>Xenopus</i> embryos, but are temporally delayed.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-08 02:07:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/767949"], "description"=>"<p>(<b>A</b>) A highly conserved putative TEAD-binding site (yellow boxes) is present in the 5′ regulatory region of the <i>pax3</i> gene in 15 different vertebrates. A previously described mouse TEAD-binding site (red box) appears less conserved. (<b>B</b>) Chromatin isolated from 300 wild type stage 14–16 <i>Xenopus laevis</i> embryos was sheared to a size range of 150 to 900 base pairs. (<b>C</b>) Chromatin immunoprecipitations (ChIPs) from 12.5 µg or 25 µg of sheared chromatin immunoprecipitated a band at the expected size for the putative novel TEAD1-binding site region with the hYAP antibody but not with a control IgG antibody. (<b>D</b>) Sequencing of this band from three different clones verified that the genomic region pulled down by the hYAP anitbody contained the novel TEAD-binding site (yellow) when compared to the <i>Xenopus tropicalis</i> genomic sequence.</p>", "links"=>[], "tags"=>["xyap", "resides"], "article_id"=>438326, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020309.g007", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Endogenous_xYAP_resides_at_a_novel_5_regulatory_region_of_pax3_/438326", "title"=>"Endogenous xYAP resides at a novel 5′ regulatory region of <i>pax3</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-08 02:18:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/767711"], "description"=>"<p>(<b>A</b>) Genes expressed in the pre-placodal ectoderm (PPE), <i>sox11</i> and <i>six1</i>, are dramatically reduced on the xYAP-injected sides (arrows). Brackets indicate the laterally located PPE expression domains on both sides the embryos. Anterior views. (<b>B</b>) Expression of the epidermis-specific <i>cyto-keratin</i> gene is lost on the xYAP-injected side. Anterior view. (<b>C</b>) The expression of genes characteristic of premigratory neural crest (<i>foxD3</i>, <i>zic1</i> at bracket) are repressed on the xYAP-injected sides. Anterior-dorsal views. (<b>D</b>) <i>pax3</i> expression in the surface ectodermal A-P stripe, which indicates the hatching gland progenitors (vertical arrows, HG) is repressed on the xYAP-injected side. In contrast, <i>pax3</i> expression in the underlying neural crest progenitors is expanded (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020309#pone-0020309-g006\" target=\"_blank\">Figure 6A</a>). Dorsal view. (<b>E</b>) <i>six1</i> expression in the otocyst (bracket) is reduced on the xYAP-injected side (right panel) compared to the uninjected side (Un) of the same embryo (left panel). Side views, stage 26. (<b>F</b>) The migratory path of <i>foxD3</i>-expressing neural crest cells (indicated by brackets) is truncated on the xYAP-injected side (right). Frontal view, stage 24. (<b>G</b>) <i>neuroD</i> expression in the trigeminal placode (bracket) is reduced on the xYAP-injected side (right panel) compared to the uninjected side (Un) of the same embryo (left panel). Side views, stage 22.</p>", "links"=>[], "tags"=>["gain-of-function", "inhibits", "genes", "pre-placodal", "premigratory", "neural", "hatching"], "article_id"=>438067, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020309.g005", "stats"=>{"downloads"=>3, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_xYAP_gain_of_function_inhibits_the_expression_of_genes_in_the_pre_placodal_ectoderm_epidermis_premigratory_neural_crest_and_hatching_gland_/438067", "title"=>"xYAP gain-of-function inhibits the expression of genes in the pre-placodal ectoderm, epidermis, premigratory neural crest, and hatching gland.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-08 02:14:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/768155"], "description"=>"<p>Sample sizes and frequencies of genes that were expanded (sox2, pax3+ neural crest progenitors [NCP]) or reduced (pax3+ hatching gland [HG] progenitors, p27xic1, sox11, myoD, and cyto-keratin) after injection of wild type xyap or mutant (defined in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020309#pone-0020309-g008\" target=\"_blank\">Fig. 8A</a>) xyap mRNAs.</p>", "links"=>[], "tags"=>["deletion", "mutants", "differential"], "article_id"=>438523, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020309.t001", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_xYAP_deletion_mutants_exhibit_differential_activities_/438523", "title"=>"xYAP deletion mutants exhibit differential activities.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-06-08 02:22:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/767582"], "description"=>"<p>(<b>A</b>) The neural plate progenitor field marked by <i>sox2</i> expression (blue stain) was darker, longer, and/or wider on the xYAP-injected side (arrow, red β-gal staining) compared to the uninjected side of the same embryo. xYAP MO-mediated knockdown (40 ng) eliminated <i>sox2</i> expression on the injected side, whereas a control MO (cMO) did not. In this and all subsequent panels: n = sample size; % = frequency of the phenotype; arrow indicates injected side. (<b>B</b>) Three genes indicative of neural differentiation (<i>neuroD</i>, <i>n-tubulin</i>, <i>p27<sup>Xic1</sup></i>) were inhibited by xYAP gain-of-function. (<b>C</b>) xYAP gain-of-function reduced <i>notch</i> and <i>hes1</i> expression. (<b>D</b>) Muscle differentiation marker, <i>myoD</i>, was reduced by xYAP gain-of-function. All views are dorsal-anterior.</p>", "links"=>[], "tags"=>["gain-of-function", "expands", "neural", "progenitor", "differentiation"], "article_id"=>437948, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020309.g004", "stats"=>{"downloads"=>2, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_xYAP_gain_of_function_expands_neural_progenitor_fields_while_neural_differentiation_is_inhibited_/437948", "title"=>"xYAP gain-of-function expands neural progenitor fields, while neural differentiation is inhibited.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-08 02:12:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/767133"], "description"=>"<p>(<b>A</b>) Frog and zebrafish YAP possess the ascribed functional and protein-protein interaction domains, including the TEAD-binding site (purple), the LATS phosphorylation site (orange), the two WW domains (red) that allow for PPxY binding, the Src Homology 3 (SH3)-binding domain (green), the coiled-coil region (blue), the transactivation domain (underline), and the PDZ-binding motif (pink). hnRNP U (yellow) binding has only been experimentally tested with human YAP, but related sites are in the fish and frog proteins. This diagram also illustrates the relative location of <i>Xenopus laevis</i> (x) and <i>Danio rerio</i> (z) MO-binding sites. (<b>B</b>) Injection of an equimolar cocktail of all three xYAP MOs at two concentrations (40 ng and 80 ng) resulted in efficient knockdown of endogenous, zygotic xYAP protein in stage 15 embryos as measured by western blot analysis. EF-2 expression from the same blot served as the loading control. (<b>C</b>) Three different xYAP MOs (80 ng; see A for binding sites) resulted in failed closure of the blastopore (arrows). (<b>D</b>) Reducing the concentration of the xYAP MO cocktail (left side) allowed blastopore closure, but resulted in dose-dependent A-P axis shortening. (<b>E</b>) Time-lapse video microscopy showed that zYAP MO (16 ng) injected embryos also exhibit perturbation in the completion of gastrulation. Asterisks mark the tissue front of epiboly movements. In uninjected and cMO-injected embryos, this front completely envelops the yolk by 10 hours post-fertilization (hpf). These fronts are still in the equatorial region in the 7.7–10 hpf YAP MO-injected embryos.</p>", "links"=>[], "tags"=>["morphant", "embryos", "defects", "a-p", "axis"], "article_id"=>437502, "categories"=>["Developmental Biology"], "users"=>["Stephen T. Gee", "Sharon L. Milgram", "Kenneth L. Kramer", "Frank L. Conlon", "Sally A. Moody"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020309.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_YAP_morphant_embryos_exhibit_defects_in_A_P_axis_elongation_/437502", "title"=>"YAP morphant embryos exhibit defects in A-P axis elongation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-08 02:05:02"}

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Relative Metric

{"start_date"=>"2011-01-01T00:00:00Z", "end_date"=>"2011-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[304, 568, 702, 818, 927, 1027, 1118, 1206, 1285, 1357, 1427, 1500, 1564, 1636, 1705, 1773, 1840, 1909, 1974, 2039, 2106, 2170, 2234, 2296, 2358, 2423, 2484, 2546, 2606, 2673, 2734, 2795, 2857, 2921, 2984, 3046, 3100]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[289, 559, 699, 817, 931, 1031, 1127, 1214, 1297, 1370, 1444, 1515, 1584, 1656, 1726, 1797, 1862, 1930, 1996, 2061, 2125, 2190, 2250, 2311, 2373, 2435, 2496, 2563, 2630, 2692, 2760, 2824, 2898, 2960, 3019, 3089, 3143]}, {"subject_area"=>"/Biology and life sciences/Neuroscience", "average_usage"=>[306, 540, 664, 768, 864, 959, 1036, 1121, 1199, 1277, 1341, 1411, 1477, 1543, 1609, 1675, 1739, 1802, 1868, 1933, 1995, 2050, 2107, 2164, 2220, 2277, 2327, 2374, 2436, 2488, 2542, 2594, 2650, 2705, 2769, 2817, 2859]}]}
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