The Unconserved Groucho Central Region Is Essential for Viability and Modulates Target Gene Specificity
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{"title"=>"The unconserved groucho central region is essential for viability and modulates target gene specificity", "type"=>"journal", "authors"=>[{"first_name"=>"Wiam", "last_name"=>"Turki-Judeh", "scopus_author_id"=>"54941298600"}, {"first_name"=>"Albert J.", "last_name"=>"Courey", "scopus_author_id"=>"7003350876"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84856517757", "pui"=>"364189192", "doi"=>"10.1371/journal.pone.0030610", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)", "sgr"=>"84856517757", "pmid"=>"22319573"}, "id"=>"d44e9ba6-6a92-30a4-9e40-9fa2f972a18f", "abstract"=>"Groucho (Gro) is a Drosophila corepressor required by numerous DNA-binding repressors, many of which are distributed in gradients and provide positional information during development. Gro contains well-conserved domains at its N- and C-termini, and a poorly conserved central region that includes the GP, CcN, and SP domains. All lethal point mutations in gro map to the conserved regions, leading to speculation that the unconserved central domains are dispensable. However, our sequence analysis suggests that the central domains are disordered leading us to suspect that the lack of lethal mutations in this region reflects a lack of order rather than an absence of essential functions. In support of this conclusion, genomic rescue experiments with Gro deletion variants demonstrate that the GP and CcN domains are required for viability. Misexpression assays using these same deletion variants show that the SP domain prevents unrestrained and promiscuous repression by Gro, while the GP and CcN domains are indispensable for repression. Deletion of the GP domain leads to loss of nuclear import, while deletion of the CcN domain leads to complete loss of repression. Changes in Gro activity levels reset the threshold concentrations at which graded repressors silence target gene expression. We conclude that co-regulators such as Gro are not simply permissive components of the repression machinery, but cooperate with graded DNA-binding factors in setting borders of gene expression. We suspect that disorder in the Gro central domains may provide the flexibility that allows this region to mediate multiple interactions required for repression.", "link"=>"http://www.mendeley.com/research/unconserved-groucho-central-region-essential-viability-modulates-target-gene-specificity", "reader_count"=>21, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>4, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>5, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>4, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>5, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>16, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>16}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"Portugal"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/349570", "https://ndownloader.figshare.com/files/349630"], "description"=>"<div><p>Groucho (Gro) is a <em>Drosophila</em> corepressor required by numerous DNA-binding repressors, many of which are distributed in gradients and provide positional information during development. Gro contains well-conserved domains at its N- and C-termini, and a poorly conserved central region that includes the GP, CcN, and SP domains. All lethal point mutations in <em>gro</em> map to the conserved regions, leading to speculation that the unconserved central domains are dispensable. However, our sequence analysis suggests that the central domains are disordered leading us to suspect that the lack of lethal mutations in this region reflects a lack of order rather than an absence of essential functions. In support of this conclusion, genomic rescue experiments with Gro deletion variants demonstrate that the GP and CcN domains are required for viability. Misexpression assays using these same deletion variants show that the SP domain prevents unrestrained and promiscuous repression by Gro, while the GP and CcN domains are indispensable for repression. Deletion of the GP domain leads to loss of nuclear import, while deletion of the CcN domain leads to complete loss of repression. Changes in Gro activity levels reset the threshold concentrations at which graded repressors silence target gene expression. We conclude that co-regulators such as Gro are not simply permissive components of the repression machinery, but cooperate with graded DNA-binding factors in setting borders of gene expression. We suspect that disorder in the Gro central domains may provide the flexibility that allows this region to mediate multiple interactions required for repression.</p> </div>", "links"=>[], "tags"=>["unconserved", "groucho", "viability", "modulates", "specificity"], "article_id"=>129105, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.s001", "https://dx.doi.org/10.1371/journal.pone.0030610.s002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/The_Unconserved_Groucho_Central_Region_Is_Essential_for_Viability_and_Modulates_Target_Gene_Specificity/129105", "title"=>"The Unconserved Groucho Central Region Is Essential for Viability and Modulates Target Gene Specificity", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-02-03 02:31:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/685218"], "description"=>"<p>Prediction of disorder in Gro using two disorder prediction algorithms: PONDR-FIT™ (Meta, black dashed line) <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030610#pone.0030610-Xue1\" target=\"_blank\">[40]</a> and FoldIndex© (green and red shaded plot) <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030610#pone.0030610-Prilusky1\" target=\"_blank\">[41]</a>. FoldIndex© tool also calculated hydrophobicity (blue line) and charge density (pink line). Residues with Meta scores exceeding 0.5 are likely to be disordered, as are residues with FoldIndex© scores below 0. The prediction tools strongly suggest that the Gro central region is disordered, while the Q and WD-repeat domains are ordered. The Gro domains (shown along the horizontal axis) are labeled with the many of the functions that have been previously ascribed to them (see text for references).</p>", "links"=>[], "tags"=>["domains", "mediate", "functions"], "article_id"=>355708, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_central_domains_mediate_multiple_essential_functions_and_are_likely_disordered_/355708", "title"=>"The central domains mediate multiple essential functions and are likely disordered.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:35:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/685286"], "description"=>"<p>(A) Structure of the Gro deletion variants, Gro<sup>ΔGP</sup>, Gro<sup>ΔCcN</sup>, Gro<sup>ΔSP</sup>, and Gro<sup>ΔCR</sup>. (B) Gro self-association assays. Untagged wild-type full-length Gro (lanes 1, 2, 7 and 8) or internal deletion variants of Gro (lanes 3–6) were translated alone (lane 1); or cotranslated with the wild-type His-tagged Gro N-terminal region (His-GroN<sup>WT</sup>, contains the first 194 amino acids of Gro including the intact Q domain) (lanes 2–6), with the 40D/89D double point mutant form of the His-tagged Gro N-terminal region (His-GroN<sup>40D/89D</sup>) (Lane 7), or the 38D/87D double point mutant form of the His-tagged Gro N-terminal region (His-GroN<sup>38D/87D</sup>) (Lane 8). Samples were incubated with Ni-NTA beads, and after extensive washing, Ni-bound proteins were eluted and analyzed by 10% SDS-PAGE and autoradiography. 10% of input is shown on the left, while the Ni-bound proteins are shown on the right. The percentage of input that was immobilized on the beads is indicated at the bottom of each lane. (C) GST-Brk pulldown assays using Gro deletion variants. [<sup>35</sup>S]-methionine-labeled Gro<sup>WT</sup> (lanes 1–3), Gro<sup>ΔGP</sup> (lanes 4–5), Gro<sup>ΔCcN</sup> (lanes 6–7), Gro<sup>ΔSP</sup> (lanes 8–9), or Gro<sup>ΔCR</sup> (lanes 10–11) were incubated with glutathione-agarose beads bearing immobilized GST (lane 2) or a GST-Brk fusion protein (lanes 3, 5, 7, 9 and 11). After extensive washing, bound proteins were eluted and analyzed by 8% SDS-PAGE and autoradiography. The percentage of input protein that bound the glutathione-agarose beads is indicated at the bottom of the GST and GST-Brk lanes.</p>", "links"=>[], "tags"=>["deletions", "impair", "wd-repeat"], "article_id"=>355778, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Central_domain_deletions_impair_neither_Q_nor_WD_repeat_domain_function_/355778", "title"=>"Central domain deletions impair neither Q nor WD-repeat domain function.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:36:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/685397"], "description"=>"<p>(A) Intron/exon organization of <i>gro</i> and flanking regions from the left arm of chromosome 3. The 10 kb region used in the rescue constructs is indicated by the bracket. Exons encoding to the Q domain are in green, GP domain in blue, CcN domain in magenta, SP domain in orange, WD repeat domain in red, and non-encoding exons in gray. (B) The hypomorphic (<i>MB12</i>) and null (<i>MB36</i>) <i>gro</i> alleles <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030610#pone.0030610-Jennings2\" target=\"_blank\">[16]</a>. The black lines indicate the sequences included in each deletion variant or mutant allele. The green lines represent the coiled-coil motifs in the Q domain that are known to be required for self-association <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030610#pone.0030610-Song1\" target=\"_blank\">[18]</a>. (C) The indicated Gro rescue constructs were tested, as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030610#s4\" target=\"_blank\">Materials and Methods</a>, for their ability to rescue the lethality associated with the MB12 or MB36 <i>gro</i> alleles. Each data point is the average (± S.D.) of three independent trials (for each of trials 1 and 2, 120 flies were analyzed; for trial 3, 100 flies were analyzed). Asterisks (*) signify p<10<sup>−5</sup> as determined from the two-tailed unpaired student's T test.</p>", "links"=>[], "tags"=>["gro", "mutant", "alleles", "genomic", "constructs", "encoding", "deletion"], "article_id"=>355882, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Rescue_of_gro_mutant_alleles_with_genomic_rescue_constructs_encoding_Gro_deletion_variants_/355882", "title"=>"Rescue of gro mutant alleles with genomic rescue constructs encoding Gro deletion variants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:38:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/685461"], "description"=>"<p>(A) qRT-PCR analysis of the <i>gro</i> transcript in embryos shows that <i>Mat-Gal4</i> driven expression results in very similar levels of overexpression of each variant. (B) An anti-Gro immunoblot verifies equal expression of the variants. An anti-tubulin immunoblot serves as a control for relative total protein levels. (C) qRT-PCR was performed on embryos lacking maternally overexpressed Gro; or containing maternally overexpressed Gro<sup>WT</sup> (green), Gro<sup>ΔGP</sup> (blue), Gro<sup>ΔCcN</sup> (pink), Gro<sup>ΔSP</sup> (orange), or Gro<sup>ΔCR</sup> (yellow). Expression levels of <i>zen</i>, <i>dpp</i>, <i>twi</i>, <i>ftz</i>, <i>hkb</i>, <i>tll</i>, <i>sna</i>, <i>kni</i>, <i>Rpt3</i>, and <i>cin</i> were normalized for expression levels of <i>RpL32</i>. Each fold repression value was obtained by dividing the normalized expression level of a gene in the absence of overexpressed Gro by the normalized expression level of the same gene in the presence of one of the overexpressed Gro variants. Values were graphed on a log<sub>2</sub> scale. Statistical significance of each value relative to the value in embryos lacking overexpressed Gro was determined from the two-tailed unpaired Student's T-test. * signifies p<0.05, ** signifies p<0.01, *** signifies p<0.005. No asterisk signifies p>0.05.</p>", "links"=>[], "tags"=>["domains", "gro-mediated", "repression"], "article_id"=>355956, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Central_domains_have_both_positive_and_negative_roles_during_Gro_mediated_repression_in_the_embryo_/355956", "title"=>"Central domains have both positive and negative roles during Gro-mediated repression in the embryo.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:39:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/685619"], "description"=>"<p>Fluorescence in situ hybridization analyzing the mRNA products of the Gro targets <i>hkb</i>, <i>tll</i>, and <i>sna</i> in cellular blastoderm embryos containing maternally overexpressed Gro variants. Embryos are oriented with the anterior to the left and ventral at the bottom. Embryo images were obtained using a 20× objective. Scale bars = 50 µm. (A–D) Wild-type expression patterns. (E–H) Gro<sup>WT</sup> overexpression leads to spatial patterning defects and reduced expression of all three target genes. Posterior <i>hkb</i> expression was absent and anterior <i>hkb</i> expression was significantly reduced (E). Posterior and anterior <i>tll</i> expression were reduced with a greater effect at the posterior (F). The ventral <i>sna</i> stripe was narrowed ventrally and retracted from both termini (G). (I–P) Overexpression of Gro<sup>ΔGP</sup> (I–L) and Gro<sup>ΔCcN</sup> (M–P) led to no changes in target gene expression patterns. (Q–T) Gro<sup>ΔSP</sup> overexpression led to defects in spatial patterning and a reduction in the expression of all three target genes that is more severe than that resulting from Gro<sup>WT</sup> overexpression. <i>hkb</i> (Q) and <i>tll</i> (R) expression were completely abolished at both termini. The <i>sna</i> stripe was substantially narrowed in the posterior region (as indicated by the arrow) and retracted from the termini, especially the anterior terminus (S). (U–X) Gro<sup>ΔCR</sup> overexpression resulted in expansion of the posterior <i>hkb</i> domain (U), and an expansion of the anterior <i>tll</i> domain toward the ventral midline of the embryo (arrow) (V). The <i>sna</i> domain is expanded towards both termini (W). Arrowheads (panels A, C, G, S, U, and W) indicate borders of the wild-type expression domains. (Y–Z) Quantification of changes in expression pattern. (Y) 20 stage 5 embryos overexpressing each variant were scored according to whether the anterior and posterior expression domains of <i>hkb</i> and <i>tll</i> appeared wild-type, were reduced, were missing altogether, or were expanded. (Z) The same 20 embryos were scored according to whether the <i>sna</i> expression domain showed no retraction, mild retraction, strong retraction, or expansion at the anterior and posterior termini.</p>", "links"=>[], "tags"=>["domains", "patterns", "embryonic", "gro"], "article_id"=>356106, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Central_domains_define_patterns_of_embryonic_Gro_target_gene_expression_/356106", "title"=>"Central domains define patterns of embryonic Gro target gene expression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:41:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/685728"], "description"=>"<p>The <i>Mat-Gal4</i> driver was used to drive maternal overexpression of the indicated Gro variants. (A–F) Representative cuticles resulting from overexpression of Gro variants. Cuticles images were obtained using a 10× objective. Scale bars = 100 µm (A) Embryos containing the driver but no UAS-Gro construct show no phenotype. Overexpression of Gro<sup>WT</sup> (B) and Gro<sup>ΔSP</sup> (E) resulted in moderate to severe cuticle defects. Overexpression of Gro<sup>ΔGP</sup> (C) resulted in no defects, while overexpression of Gro<sup>ΔCcN</sup> (D) resulted in a truncated or missing 4<sup>th</sup> or 6<sup>th</sup> abdominal denticle belt (indicted by arrow), a phenotype sometimes observed in weak <i>gro</i> hypomorphic embryos, and thus consistent with a weak dominant negative function for this deletion variant. Overexpression of Gro<sup>ΔCR</sup> (F) often resulted in a pair-rule defect reminiscent of that seen in <i>eve</i> mutant embryos. This is consistent with the notion that Gro<sup>ΔCR</sup> is a dominant negative, since Gro is required for Eve function <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030610#pone.0030610-Mannervik1\" target=\"_blank\">[35]</a>, <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030610#pone.0030610-Kobayashi1\" target=\"_blank\">[61]</a>. (G) Cuticles of 100 embryos laid by females overexpressing each of the five Gro variants were assigned to the following phenotypic categories: Wild-type - no observable defects; Minor defects - 1–2 missing or fused denticle belts; Moderate defects - 3–4 missing and/or fused denticle belts; Severe defects - 5 or more missing denticle belts. (H) 100 embryos overexpressing each variant were scored to determine percent of embryos that deposited cuticle and that hatched.</p>", "links"=>[], "tags"=>["phenotypes", "resulting", "overexpression", "gro"], "article_id"=>356220, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cuticle_phenotypes_resulting_from_overexpression_of_Gro_variants_/356220", "title"=>"Cuticle phenotypes resulting from overexpression of Gro variants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:43:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/685814"], "description"=>"<p>Clones of Gro variant overexpressing cells were generated in third instar wing discs contained the <i>vgQ-lacZ</i> reporter. Discs were stained with antibodies against CD2 (green) and β-galactosidase (red). Overexpression clones are marked by the absence of CD2. Arrows mark clones that overlap the <i>vg</i> expression domain. Gro<sup>WT</sup> clones (B) and Gro<sup>ΔSP</sup> clones (E), exhibited ectopic repression of <i>vgQ-lacZ</i>, while control clones (A), Gro<sup>ΔGP</sup> clones (C), and Gro<sup>ΔCcN</sup> clones (D) exhibited no repression of <i>vgQ-lacZ</i>. Gro<sup>ΔCR</sup> clones (F) exhibited slight expansion of <i>vgQ-lacZ</i> into regions in which the reporter is not normally expressed. Clones containing a mixture of overexpressed Gro<sup>WT</sup> and Gro<sup>ΔCR</sup> (G) show significantly reduced ectopic repression of <i>vgQ-lacZ</i>, relative to clones overexpressing Gro<sup>WT</sup> alone. Wing disc images were obtained using a 20× objective. Scale bars = 50 µm.</p>", "links"=>[], "tags"=>["domains", "gro-mediated", "repression"], "article_id"=>356304, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Central_domains_regulate_Gro_mediated_repression_in_the_wing_disc_/356304", "title"=>"Central domains regulate Gro-mediated repression in the wing disc.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:45:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/685931"], "description"=>"<p>The <i>Ser-Gal4</i> driver was used to drive expression of Gro<sup>WT</sup> or Gro internal deletion variants in the wing. (A) A wing containing the driver but no UAS-Gro construct shows no phenotype. Overexpression of Gro<sup>WT</sup> (B) and Gro<sup>ΔSP</sup> (E) resulted in moderate to severe blistering and vein deformation. Overexpression of Gro<sup>ΔGP</sup> (C) resulted in no defects, while overexpression of Gro<sup>ΔCcN</sup> (D) resulted in bifurcation of the 4<sup>th</sup> and/or 5<sup>th</sup> longitudinal vein (LV, arrows). Overexpression of Gro<sup>ΔCR</sup> (F) resulted in severe blistering and vein deformation as well as ectopic bristles along the wing vein (inset, yellow arrows). (G) 100 wings of each genotype were scored according to phenotype as indicated showing the differences in the relative severity of the phenotypes generated by overexpression of the various deletion variants. (H) Gro immunoblot of third instar wing discs verifies equal expression levels. Tubulin serves as a control for equal protein levels. (I) In addition to the blistering and wing veination defects, Gro<sup>ΔCcN</sup> and Gro<sup>ΔCR</sup> Gro overexpression also resulted in 13% and 29% wing scalloping phenotypes (arrows). The severe defects in the Gro<sup>ΔCR</sup> overexpressing wings and the milder defects in the Gro<sup>ΔCcN</sup> overexpressing wings probably results from a dominant negative function for these variant as discussed in the text. The wing scalloping (J) and the ectopic wing bristles (F and I, indicated by arrows in the high magnification insets), both of which are reminiscent of a Notch pathway hypomorphic phenotypes <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030610#pone.0030610-Hartenstein1\" target=\"_blank\">[49]</a>. Except for the high magnification insets in panels F and I, the wing images were obtained using a 4× objective. Scale bars = 400 µm. For the high magnification insets in panels F and I, the images were obtained using a 40× objective.</p>", "links"=>[], "tags"=>["resulting", "overexpression", "gro", "variants"], "article_id"=>356423, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phenotypes_resulting_from_overexpression_of_Gro_variants_in_the_wing_disc_/356423", "title"=>"Phenotypes resulting from overexpression of Gro variants in the wing disc.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:47:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/686028"], "description"=>"<p>(A–E) Third instar imaginal wing discs were stained with Myc antibodies (red) to detect Myc-tagged Gro and DAPI (blue) to stain DNA. Gro<sup>WT</sup> (A) and Gro<sup>ΔSP</sup> (D) localized exclusively to the nucleus. Gro<sup>ΔGP</sup> (B) localized exclusively to the cytoplasm. Gro<sup>ΔCcN</sup> (C) localized primarily to the nucleus, but we also detected a low level of cytoplasmic localization and Gro<sup>ΔCR</sup> (E) localized to the nucleus and cytoplasm. Wing disc images were obtained using a 100× objective. Scale bars = 10 µm. (F–J) <i>Drosophila</i> S2 cells were stained with FLAG antibodies (red) to detect FLAG-tagged Gro and DAPI (blue) to stain DNA. Gro<sup>WT</sup> (F) and Gro<sup>ΔSP</sup> (I) localized exclusively to the nucleus. Gro<sup>ΔGP</sup> (G) and Gro<sup>ΔCR</sup> (J) localized exclusively to the cytoplasm. Gro<sup>ΔCcN</sup> (H) localized primarily to the nucleus. S2 cell images were obtained using a 100× objective. Scale bars = 5 µm.</p>", "links"=>[], "tags"=>["localization", "gro", "deletion"], "article_id"=>356517, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g009"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Subcellular_localization_of_Gro_central_domain_deletion_variants_/356517", "title"=>"Subcellular localization of Gro central domain deletion variants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:48:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/686113"], "description"=>"<p>(A) GST-tagged histone H3 and H4 tails were expressed in bacteria and purified on glutathione beads. 750 ng of GST-H3 (lane 1), GST-H4 (lane 2), and GST (lane 3) were subjected to 10% SDS-PAGE followed by staining with Coomassie Blue. Lane 4 shows size markers with molecular mass indicated in kDa. (B) Gro<sup>WT</sup> (lane 1), Gro<sup>ΔGP</sup> (lane 2), Gro<sup>ΔCcN</sup> (lane 3), Gro<sup>ΔSP</sup> (lane 4), and Gro<sup>ΔCR</sup> (lane 5) were translated in vitro in the presence of [<sup>35</sup>S] methionine. 10% of the translation products were subjected to 8% SDS-PAGE and the gel imaged by autoradiography. (C) The translation products from (B) were incubated with glutathione bead bound GST-H3 (Lanes 1–5), GST-H4 (Lanes 6–10), or GST (Lane 11). After extensive washing, bound proteins were eluted with SDS gel sample buffer and subjected to 8% SDS-PAGE. The gel was imaged by autoradiography. The percentage of input protein that bound the immobilized GST fusion proteins is indicated at the bottom of each lane.</p>", "links"=>[], "tags"=>["contains", "redundant", "determinants", "binding", "histone", "h3", "h4"], "article_id"=>356601, "categories"=>["Biological Sciences", "Biochemistry", "Genetics", "Developmental Biology"], "users"=>["Wiam Turki-Judeh", "Albert J. Courey"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030610.g010"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Central_region_contains_redundant_determinants_of_binding_to_the_histone_H3_and_H4_tails_/356601", "title"=>"Central region contains redundant determinants of binding to the histone H3 and H4 tails.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-03 01:50:01"}

PMC Usage Stats | Further Information

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Relative Metric

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