Functional Study of Mammalian Neph Proteins in Drosophila melanogaster
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{"title"=>"Functional study of mammalian Neph proteins in Drosophila melanogaster", "type"=>"journal", "authors"=>[{"first_name"=>"Martin", "last_name"=>"Helmstädter", "scopus_author_id"=>"45860935400"}, {"first_name"=>"Kevin", "last_name"=>"Lüthy", "scopus_author_id"=>"57200225081"}, {"first_name"=>"Markus", "last_name"=>"Gödel", "scopus_author_id"=>"13405898700"}, {"first_name"=>"Matias", "last_name"=>"Simons", "scopus_author_id"=>"7201538823"}, {"last_name"=>"Ashish", "scopus_author_id"=>"24774275700"}, {"first_name"=>"Deepak", "last_name"=>"Nihalani", "scopus_author_id"=>"6506082766"}, {"first_name"=>"Stefan A.", "last_name"=>"Rensing", "scopus_author_id"=>"6603747563"}, {"first_name"=>"Karl Friedrich", "last_name"=>"Fischbach", "scopus_author_id"=>"7004238918"}, {"first_name"=>"Tobias B.", "last_name"=>"Huber", "scopus_author_id"=>"7103351974"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "sgr"=>"84863688902", "scopus"=>"2-s2.0-84863688902", "pui"=>"365220930", "doi"=>"10.1371/journal.pone.0040300", "pmid"=>"22792268"}, "id"=>"d51e3562-2c32-39e8-8306-661c0e8ae222", "abstract"=>"Neph molecules are highly conserved immunoglobulin superfamily proteins (IgSF) which are essential for multiple morphogenetic processes, including glomerular development in mammals and neuronal as well as nephrocyte development in D. melanogaster. While D. melanogaster expresses two Neph-like proteins (Kirre and IrreC/Rst), three Neph proteins (Neph1-3) are expressed in the mammalian system. However, although these molecules are highly abundant, their molecular functions are still poorly understood. Here we report on a fly system in which we overexpress and replace endogenous Neph homologs with mammalian Neph1-3 proteins to identify functional Neph protein networks required for neuronal and nephrocyte development. Misexpression of Neph1, but neither Neph2 nor Neph3, phenocopies the overexpression of endogenous Neph molecules suggesting a functional diversity of mammalian Neph family proteins. Moreover, structure-function analysis identified a conserved and specific Neph1 protein motif that appears to be required for the functional replacement of Kirre. Hereby, we establish D. melanogaster as a genetic system to specifically model molecular Neph1 functions in vivo and identify a conserved amino acid motif linking Neph1 to Drosophila Kirre function.", "link"=>"http://www.mendeley.com/research/functional-study-mammalian-neph-proteins-drosophila-melanogaster", "reader_count"=>22, "reader_count_by_academic_status"=>{"Student > Doctoral Student"=>4, "Researcher"=>2, "Student > Ph. D. Student"=>9, "Student > Master"=>3, "Student > Bachelor"=>4}, "reader_count_by_user_role"=>{"Student > Doctoral Student"=>4, "Researcher"=>2, "Student > Ph. D. Student"=>9, "Student > Master"=>3, "Student > Bachelor"=>4}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>7, "Agricultural and Biological Sciences"=>11, "Medicine and Dentistry"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>11}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>7}}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/615023"], "description"=>"<p><b>A.</b> Scanning electron micrographs of GCNs at third larval stage. Scale bar: 20 µm. <b>B.</b> The fact that hyperfused GCNs lose their spherical shape was used to quantify the rescue efficiency. Kirre or Neph1 expression is sufficient to significantly rescue the <i>kirre<sup>-</sup></i> phenotype. *P value <0,0001 (unpaired t-test with Welchs correction). Genotypes: Control: <i>+/sns-GAL4. kirre<sup>-</sup></i>: <i>Df(1)duf</i> <i><sup>sps-1</sup>/y</i>. Kirre rescue: <i>Df(1)duf</i> <i><sup>sps-1</sup>/y; UAS-Kirre/sns-GAL4.</i> Neph1 rescue: <i>Df(1)duf</i> <i><sup>sps-1</sup>/y; UAS-neph1/sns-GAL4</i>. Neph2 rescue: <i>Df(1)duf</i> <i><sup>sps-1</sup>/y; UAS-neph2/sns-GAL4</i>. Neph3 rescue: <i>Df(1)duf</i> <i><sup>sps-1</sup>/y; UAS-neph3/sns-GAL4.</i></p>", "links"=>[], "tags"=>["larval", "gcns", "neph1"], "article_id"=>285499, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Marine Biology"], "users"=>["Martin Helmstädter", "Kevin Lüthy", "Markus Gödel", "Matias Simons", "Ashish", "Deepak Nihalani", "Stefan A. Rensing", "Karl-Friedrich Fischbach", "Tobias B. Huber"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040300.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Third_larval_stage_kirre_GCNs_are_hyperfused_Neph1_can_rescue_the_kirre_phenotype_/285499", "title"=>"Third larval stage <i>kirre<sup>-</sup></i> GCNs are hyperfused. Neph1 can rescue the <i>kirre<sup>-</sup></i> phenotype.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-06 01:31:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/615458"], "description"=>"<p>Scanning electron micrograph of adult <i>Drosophila</i> eyes (<b>A,B,F,G,K,L,P,Q</b>), close-up of the eye (<b>B,G,L,Q</b>) and light micrographs of semithin sections (<b>C,H,M,R</b>). The control shows the regular crystal like arrangement of an <i>Drosophila</i> eye (<b>A,B</b>). <i>Sev-GAL4</i> induced misexpression of IrreC/Rst (<b>F,G</b>) or Kirre (<b>K,L</b>) results in a rough eye phenotype. Misexpression of Neph1 with <i>sev-GAL4</i> also causes a rough eye phenotype (<b>P,Q</b>). All three genotypes exhibit fusion of ommatidia (<b>H,M,R</b>). Genotypes: <i>sev-GAL4/+</i> (<b>A,B,C</b>); <i>sev-GAL4/UAS-irreC/rst</i> (<b>F,G,H</b>); <i>sev-GAL4/+,UAS-kirre/+</i> (<b>K,L</b>); <i>sev-GAL4/UAS-neph1_V5</i> (<b>P,Q,R</b>). Auto fluorescence micrographs of adult <i>Drosophila</i> optic lobes (<b>D,I,N,S</b>). The control fly shows the typical wildtype-like arrangement of the neuropils of the <i>Drosophila</i> optic lobe (<b>D,E</b>). Overexpression of Rst (<b>I</b>) or Kirre (<b>N</b>) causes severe misrouting of fibers in medulla and lobula complex and a disorganization of these neuropils (<b>J,O</b>). Misexpression of Neph1 leads to a similar phenotype (<b>S,T</b>). Optic lobe drawing: la: lamina, me: medulla, lo: lobula, lp: lobula plate. Genotypes: <i>Mz1369-GAL4/+</i> (<b>D</b>); <i>Mz1369-GAL4/UAS-rst</i> (<b>I</b>); <i>Mz1369-GAL4/+,UAS-kirre/+</i> (<b>N</b>); <i>Mz1369-GAL4/UAS-neph1_V5</i> (<b>S</b>).</p>", "links"=>[], "tags"=>["mimic", "neuronal", "phenotypes", "overexpressed", "kirre"], "article_id"=>285940, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Marine Biology"], "users"=>["Martin Helmstädter", "Kevin Lüthy", "Markus Gödel", "Matias Simons", "Ashish", "Deepak Nihalani", "Stefan A. Rensing", "Karl-Friedrich Fischbach", "Tobias B. Huber"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040300.g006", "stats"=>{"downloads"=>1, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Neph1_can_mimic_neuronal_and_eye_phenotypes_of_overexpressed_Kirre_or_IrreC_Rst_/285940", "title"=>"Neph1 can mimic neuronal and eye phenotypes of overexpressed Kirre or IrreC/Rst.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-06 01:39:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/614849"], "description"=>"<p><b>A-F.</b> Scanning electron micrograph of <i>Drosophila</i> GCNs at third larval stage and immunoreactivity of the corresponding genotypes. The control shows the distribution of misexpressed membrane associated mCD8::GFP. The misexpressed protein is endocytosed if it is not stabilized in the nephrocyte diaphragm. The binucleate GCNs are separated (<b>A</b>). IrreC/Rst misexpression leads to clustering and hyperfusion of GCNs (<b>B</b>). Kirre overexpression leads to a similar phenotype. (<b>C</b>). Neph1 misexpression also leads to clustering and fusion of GCNs (<b>D</b>)<b>.</b> Neph2 misexpression does not interfere with the fusion of GCNs (<b>E</b>). The arrow head marks the enriched Neph2 immunoreactivity at cell-cell contacts. Misexpression of Neph3 does not interfere with the GCN fusion. Immunoreactivity shows that the Neph3 expression pattern is similar to the GFP control (<b>F</b>).</p>", "links"=>[], "tags"=>["misexpression", "gcns", "induce", "clustering"], "article_id"=>285327, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Marine Biology"], "users"=>["Martin Helmstädter", "Kevin Lüthy", "Markus Gödel", "Matias Simons", "Ashish", "Deepak Nihalani", "Stefan A. Rensing", "Karl-Friedrich Fischbach", "Tobias B. Huber"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040300.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_IRM_protein_misexpression_in_GCNs_can_induce_clustering_and_or_hyperfusion_/285327", "title"=>"IRM protein misexpression in GCNs can induce clustering and/or hyperfusion.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-06 01:28:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/614701"], "description"=>"<p><b>A.</b> Similarity network of Neph1–3, Kirre and IrreC/Rst generated with T-Coffee multiple sequence alignment algorithm. Values represent full length protein similarity in percent. <b>B.</b> Protein tree of the Neph (blue) and Nephrin (red) protein families as present in <i>Mus musculus</i>, <i>Danio rerio</i>, <i>Caenorhabditis elegans, Anopheles gambiae</i> and several <i>Drosophila</i> species <i>(Drosophila mojavensis</i> (DROMO), <i>Drosophila virilis</i> (DROVI), <i>Drosophila grimshawi</i> (DROGR), <i>Drosophila willistoni</i> (DROWI), <i>Drosophila pseudoobscura</i> (DROPS), <i>Drosophila persimilis</i> (DROPE), <i>Drosophila ananassae</i> (DROAN), <i>Drosophila melanogaster</i> (DROME), <i>Drosophila sechellia</i> (DROSE), <i>Drosophila yakuba</i> (DROYA). Arrowheads mark the proteins investigated in this study. <b>C.</b> Structural comparison of <i>M. musculus</i> kidney glomerulus (<b>e</b>), podocytes (<b>f</b>) and slit diaphragm (<b>g</b>) with GCNs of <i>D. melanogaster</i> (<b>a</b>). The rough surface of the GCN underneath the basement membrane (<b>b</b>) is formed by the nephrocyte diaphragm (<b>c</b>). In contrast to the mammalian slit diaphragm which is formed between neighboring podocytes (<b>h</b>), the <i>Drosophila</i> nephrocyte diaphragm is formed within one GCN (<b>d</b>).</p>", "links"=>[], "tags"=>["irre", "module", "conserved"], "article_id"=>285181, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Marine Biology"], "users"=>["Martin Helmstädter", "Kevin Lüthy", "Markus Gödel", "Matias Simons", "Ashish", "Deepak Nihalani", "Stefan A. Rensing", "Karl-Friedrich Fischbach", "Tobias B. Huber"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040300.g001", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_irre_cell_recognition_module_IRM_is_conserved_across_species_/285181", "title"=>"The irre cell recognition module (IRM) is conserved across species.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-06 01:26:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/615263"], "description"=>"<p><b>A.</b> Scanning electron micrographs of GCNs at third larval stage. Scale bar: 20 µm. <b>B.</b> Quantitation of GCN circularity revealing that Kirre versions CT1– CT3 which still contain the KIN1 motif are able to partially restore the wildtype situation. However, Kirre-CT4 missing the conserved KIN1 motif is not able to rescue the GCN fusion phenotype. *P value <0,0001 (unpaired t-test with Welchs correction). Genotypes: Control: <i>+/sns-GAL4</i>; <i>kirre<sup>-</sup></i>: <i>Df(1)duf</i> <i><sup>sps-1</sup></i>; CT1 rescue: <i>Df(1)duf</i> <i><sup>sps-1</sup>/y; UAS-CT1/sns-GAL4</i>; CT2 rescue: <i>Df(1)duf</i> <i><sup>sps-1</sup>/y; UAS-CT2/sns-GAL4</i>; CT3 rescue: <i>Df(1)duf</i> <i><sup>sps-1</sup>/y; UAS-CT3/sns-GAL4,</i> CT4 rescue: <i>Df(1)duf</i> <i><sup>sps-1</sup>/y; UAS-CT4/sns-GAL4.</i></p>", "links"=>[], "tags"=>["fusion", "requires", "cytoplasmic", "kirre", "containing", "kin1"], "article_id"=>285739, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Marine Biology"], "users"=>["Martin Helmstädter", "Kevin Lüthy", "Markus Gödel", "Matias Simons", "Ashish", "Deepak Nihalani", "Stefan A. Rensing", "Karl-Friedrich Fischbach", "Tobias B. Huber"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040300.g005", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GCN_fusion_requires_the_cytoplasmic_part_of_Kirre_containing_the_KIN1_motif_/285739", "title"=>"GCN fusion requires the cytoplasmic part of Kirre containing the KIN1 motif.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-06 01:35:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/319030", "https://ndownloader.figshare.com/files/319067", "https://ndownloader.figshare.com/files/319105"], "description"=>"<div><p>Neph molecules are highly conserved immunoglobulin superfamily proteins (IgSF) which are essential for multiple morphogenetic processes, including glomerular development in mammals and neuronal as well as nephrocyte development in <em>D. melanogaster</em>. While <em>D. melanogaster</em> expresses two Neph-like proteins (Kirre and IrreC/Rst), three Neph proteins (Neph1–3) are expressed in the mammalian system. However, although these molecules are highly abundant, their molecular functions are still poorly understood. Here we report on a fly system in which we overexpress and replace endogenous Neph homologs with mammalian Neph1–3 proteins to identify functional Neph protein networks required for neuronal and nephrocyte development. Misexpression of Neph1, but neither Neph2 nor Neph3, phenocopies the overexpression of endogenous Neph molecules suggesting a functional diversity of mammalian Neph family proteins. Moreover, structure-function analysis identified a conserved and specific Neph1 protein motif that appears to be required for the functional replacement of Kirre. Hereby, we establish <em>D. melanogaster</em> as a genetic system to specifically model molecular Neph1 functions <em>in vivo</em> and identify a conserved amino acid motif linking Neph1 to <em>Drosophila</em> Kirre function.</p> </div>", "links"=>[], "tags"=>["mammalian", "neph", "proteins"], "article_id"=>122969, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Marine Biology"], "users"=>["Martin Helmstädter", "Kevin Lüthy", "Markus Gödel", "Matias Simons", "Ashish", "Deepak Nihalani", "Stefan A. Rensing", "Karl-Friedrich Fischbach", "Tobias B. Huber"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0040300.s001", "https://dx.doi.org/10.1371/journal.pone.0040300.s002", "https://dx.doi.org/10.1371/journal.pone.0040300.s003"], "stats"=>{"downloads"=>3, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Functional_Study_of_Mammalian_Neph_Proteins_in_Drosophila_melanogaster_/122969", "title"=>"Functional Study of Mammalian Neph Proteins in <em>Drosophila melanogaster</em>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-07-06 00:49:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/615147"], "description"=>"<p><b>A.</b> MEME generated Bitlogo of the KIN1 motif. <b>B.</b> Structural comparison of Neph and Neph-like proteins. Blue: Ig domain. Grey: transmembrane domain. Yellow: KIN1 motif. Scale: number of amino acids. <b>C.</b> Surface representation of the structural model of the cytoplasmic domain of Neph1 is shown in green with the KIN1 motif highlighted in yellow.</p>", "links"=>[], "tags"=>["kin1", "motif"], "article_id"=>285623, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Marine Biology"], "users"=>["Martin Helmstädter", "Kevin Lüthy", "Markus Gödel", "Matias Simons", "Ashish", "Deepak Nihalani", "Stefan A. Rensing", "Karl-Friedrich Fischbach", "Tobias B. Huber"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040300.g004", "stats"=>{"downloads"=>1, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_drawing_of_the_identified_KIN1_motif_and_its_position_in_the_protein_sequences_/285623", "title"=>"Schematic drawing of the identified KIN1 motif and its position in the protein sequences.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-06 01:33:43"}

PMC Usage Stats | Further Information

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