CRIS—A Novel cAMP-Binding Protein Controlling Spermiogenesis and the Development of Flagellar Bending
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{"title"=>"CRIS-A Novel cAMP-Binding Protein Controlling Spermiogenesis and the Development of Flagellar Bending", "type"=>"journal", "authors"=>[{"first_name"=>"Anke Miriam", "last_name"=>"Krähling", "scopus_author_id"=>"56010554500"}, {"first_name"=>"Luis", "last_name"=>"Alvarez", "scopus_author_id"=>"56978120600"}, {"first_name"=>"Katharina", "last_name"=>"Debowski", "scopus_author_id"=>"55214184100"}, {"first_name"=>"Qui", "last_name"=>"Van", "scopus_author_id"=>"8554788600"}, {"first_name"=>"Monika", "last_name"=>"Gunkel", "scopus_author_id"=>"56010460000"}, {"first_name"=>"Stephan", "last_name"=>"Irsen", "scopus_author_id"=>"10043834600"}, {"first_name"=>"Ashraf", "last_name"=>"Al-Amoudi", "scopus_author_id"=>"6602578934"}, {"first_name"=>"Timo", "last_name"=>"Strünker", "scopus_author_id"=>"15756316500"}, {"first_name"=>"Elisabeth", "last_name"=>"Kremmer", "scopus_author_id"=>"24179132500"}, {"first_name"=>"Eberhard", "last_name"=>"Krause", "scopus_author_id"=>"7102585503"}, {"first_name"=>"Ingo", "last_name"=>"Voigt", "scopus_author_id"=>"16507799400"}, {"first_name"=>"Simone", "last_name"=>"Wörtge", "scopus_author_id"=>"36505258100"}, {"first_name"=>"Ari", "last_name"=>"Waisman", "scopus_author_id"=>"6701659239"}, {"first_name"=>"Ingo", "last_name"=>"Weyand", "scopus_author_id"=>"6603568018"}, {"first_name"=>"Reinhard", "last_name"=>"Seifert", "scopus_author_id"=>"8752397600"}, {"first_name"=>"Ulrich Benjamin", "last_name"=>"Kaupp", "scopus_author_id"=>"7005161976"}, {"first_name"=>"Dagmar", "last_name"=>"Wachten", "scopus_author_id"=>"55545956600"}], "year"=>2013, "source"=>"PLoS Genetics", "identifiers"=>{"isbn"=>"1553-7390 (Print)", "pmid"=>"24339785", "doi"=>"10.1371/journal.pgen.1003960", "pui"=>"372170310", "issn"=>"15537390", "sgr"=>"84892751544", "scopus"=>"2-s2.0-84892751544"}, "id"=>"7202b761-b8ce-32f6-bd38-222eb329938e", "abstract"=>"The second messengers cAMP and cGMP activate their target proteins by binding to a conserved cyclic nucleotide-binding domain (CNBD). Here, we identify and characterize an entirely novel CNBD-containing protein called CRIS (cyclic nucleotide receptor involved in sperm function) that is unrelated to any of the other members of this protein family. CRIS is exclusively expressed in sperm precursor cells. Cris-deficient male mice are either infertile due to a lack of sperm resulting from spermatogenic arrest, or subfertile due to impaired sperm motility. The motility defect is caused by altered Ca(2+) regulation of flagellar beat asymmetry, leading to a beating pattern that is reminiscent of sperm hyperactivation. Our results suggest that CRIS interacts during spermiogenesis with Ca(2+)-regulated proteins that--in mature sperm--are involved in flagellar bending.", "link"=>"http://www.mendeley.com/research/crisa-novel-campbinding-protein-controlling-spermiogenesis-development-flagellar-bending", "reader_count"=>23, "reader_count_by_academic_status"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>4, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>1, "Student > Postgraduate"=>3, "Student > Master"=>4, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>4, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>1, "Student > Postgraduate"=>3, "Student > Master"=>4, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>13, "Medicine and Dentistry"=>1, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>13}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>4}}, "reader_count_by_country"=>{"Mexico"=>2, "Germany"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1305675", "https://ndownloader.figshare.com/files/1305676", "https://ndownloader.figshare.com/files/1305677", "https://ndownloader.figshare.com/files/1305678", "https://ndownloader.figshare.com/files/1305679"], "description"=>"<div><p>The second messengers cAMP and cGMP activate their target proteins by binding to a conserved <u>c</u>yclic <u>n</u>ucleotide-<u>b</u>inding <u>d</u>omain (CNBD). Here, we identify and characterize an entirely novel CNBD-containing protein called CRIS (<u>c</u>yclic nucleotide <u>r</u>eceptor <u>i</u>nvolved in <u>s</u>perm function) that is unrelated to any of the other members of this protein family. CRIS is exclusively expressed in sperm precursor cells. <i>Cris</i>-deficient male mice are either infertile due to a lack of sperm resulting from spermatogenic arrest, or subfertile due to impaired sperm motility. The motility defect is caused by altered Ca<sup>2+</sup> regulation of flagellar beat asymmetry, leading to a beating pattern that is reminiscent of sperm hyperactivation. Our results suggest that CRIS interacts during spermiogenesis with Ca<sup>2+</sup>-regulated proteins that—in mature sperm—are involved in flagellar bending.</p></div>", "links"=>[], "tags"=>["camp-binding", "controlling", "spermiogenesis", "flagellar"], "article_id"=>870220, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>["https://dx.doi.org/10.1371/journal.pgen.1003960.s001", "https://dx.doi.org/10.1371/journal.pgen.1003960.s002", "https://dx.doi.org/10.1371/journal.pgen.1003960.s003", "https://dx.doi.org/10.1371/journal.pgen.1003960.s004", "https://dx.doi.org/10.1371/journal.pgen.1003960.s005"], "stats"=>{"downloads"=>4, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CRIS_8212_A_Novel_cAMP_Binding_Protein_Controlling_Spermiogenesis_and_the_Development_of_Flagellar_Bending_/870220", "title"=>"CRIS—A Novel cAMP-Binding Protein Controlling Spermiogenesis and the Development of Flagellar Bending", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305636"], "description"=>"<p>(<b>A</b>) Testis, epididymis, and body weight of wild-type (+/+) and knockout mice (−/−). (<b>B</b>) Analysis of testis morphology using Periodic acid-Schiff staining (PAS) of frozen testis sections. Left: wild-type testis, middle: knockout (−/−) testis with normal testis weight, right: knockout (−/−) testis with lower testis weight. Scale bars are indicated. (<b>C</b>) Analysis of tubule diameter wild-type (+/+) and knockout mice (−/−) with either normal testis weight (subfertile) or lower testis weight (infertile). Number of tubules are indicated. (<b>D–E</b>) DNA-content analysis of male germ cells. (<b>D</b>) Representative histogram of propidium iodide-stained wild-type (+/+, top) and knockout (−/−, bottom, with lower testis weight) male germ cells analyzed by FACS. (<b>E</b>) Average data (mean ± s.d) of DNA content analysis from wild-type (+/+) and knockout (−/−) male germ cells with normal testis weight. Number of animals are indicated. (<b>F</b>) Immunohistochemical analysis of MIWI in mouse testis. Testis sections (+/+: wild-type, −/−: CRIS knockout with normal testis weight) have been probed with a polyclonal MIWI-specific antibody and a fluorescent secondary antibody (red). DNA was stained with DAPI (blue). Pictures on the right show a higher magnification of a single tubule. Scale bars are indicated. (<b>G</b>) Breeding performance of wild-type (left, blue) and knockout males (right, black) mated with wild-type females. Days between first mating and birth of litter are plotted. The number of litters per day is expressed as a percentage of the total number of litters.</p>", "links"=>[], "tags"=>["mice", "prone", "spermatogenic"], "article_id"=>870185, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.g004", "stats"=>{"downloads"=>2, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cris_deficient_mice_are_prone_to_spermatogenic_arrest_/870185", "title"=>"<i>Cris</i>-deficient mice are prone to spermatogenic arrest.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305630"], "description"=>"<p>(<b>A</b>) Analysis of <i>Cris</i> mRNA expression by Northern blot. Left, mouse multi-tissue; right, mouse reproductive tissue. (<b>B</b>) Analysis of CRIS protein expression by immunoblotting using a CRIS-specific polyclonal antibody. Protein lysates: T, testis (50 µg); G, germ cells (50 µg); S, cauda sperm (100 µg); HA, mCRIS-HA expressing HEK293 cells (15 µg). Loading control: β-tubulin. (<b>C</b>) Identification of mCRIS in testis using mass spectrometry. Testis lysates were separated on a 1D SDS-PAGE, lanes were sliced, and analyzed by mass spectrometry. Unique peptides for mCRIS are indicated in blue. (<b>D</b>) Developmental expression pattern of mCRIS in testis. Proteins from mouse testis (30 µg/lane) have been probed with a CRIS-specific monoclonal antibody. The age of the mice (days after birth) is indicated. Control: mCRIS-HA expressing HEK cells (10 µg/lane); loading control: β-actin. (<b>E</b>) <i>In situ</i> hybridization. Testis sections (+/+: wild-type, −/−: CRIS knockout) have been labeled with a <i>Cris</i>-specific anti-sense probe. Dark staining indicates a positive signal. The corresponding sense probe showed no staining. Scale bars are indicated. (<b>F</b>) Immunohistochemical analysis of CRIS in mouse testis. Testis sections (+/+: wild-type, −/−: CRIS knockout) have been probed with a polyclonal CRIS-specific antibody and a fluorescent secondary antibody (red). DNA was stained with DAPI (blue). The secondary antibody unspecifically labels the interstitial cells in between the tubules (see −/−, right, secondary antibody only). Scale bars are indicated. (<b>G</b>) See (F) Higher magnification; dotted line: <i>lamina propria</i>. (<b>H</b>) Targeting strategy for the generation of <i>Cris</i>-deficient mice. Exons 5–7 (yellow boxes) have been replaced with a neomycin cassette (Neo) flanked by two lox-P elements (red arrow heads). Restriction sites, the corresponding fragment sizes, and the localization of probe 1 are indicated. (<b>I–J</b>) Verification of gene targeting. (<b>I</b>) Southern blot analysis using probe 1. The sizes of the fragments are indicated. (<b>J</b>) Analysis of CRIS expression in germ cells using a polyclonal CRIS-specific antibody (50 µg/lane). Loading control: calnexin.</p>", "links"=>[], "tags"=>["sperm"], "article_id"=>870179, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.g003", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CRIS_is_exclusively_expressed_in_sperm_precursor_cells_/870179", "title"=>"CRIS is exclusively expressed in sperm precursor-cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305651"], "description"=>"<p>IB, immunoblotting; IF, immunofluorescence.</p>", "links"=>[], "tags"=>[], "article_id"=>870196, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.t004", "stats"=>{"downloads"=>2, "page_views"=>36, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Antibodies_and_their_dilutions_/870196", "title"=>"Antibodies and their dilutions.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305648"], "description"=>"<p>Primer sequences.</p>", "links"=>[], "tags"=>[], "article_id"=>870193, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.t003", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Primer_sequences_/870193", "title"=>"Primer sequences.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305647"], "description"=>"<p>(<b>A</b>) Analysis of protein expression in testis from wild-type (+/+) and <i>Cris</i>-deficient mice (−/−) by immunoblotting. Per lane, 50 µg total testis lysates have been loaded. Antibodies against KIF2A, IFT172, and ABCF2 are described in <a href=\"http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003960#pgen-1003960-t004\" target=\"_blank\">Table 4</a>. Loading control: β-tubulin. (<b>B</b>) Immunohistochemical analysis of IFT172 and KIF2A in mouse testis. Testis sections (+/+: wild-type, −/−: CRIS knockout) have been probed with IFT2- and KIF2A-specific antibodies and a fluorescent secondary antibody (green). DNA was stained with DAPI (blue). Scale bars are indicated. (<b>C</b>) Analysis of protein expression in sperm from wild-type (+/+) and <i>Cris</i>-deficient mice (−/−) by immunoblotting. Per lane, total protein from 5×10<sup>6</sup> cells has been loaded. Loading control: β-tubulin. (<b>D</b>) Immunocytochemical analysis of IFT172 and KIF2A in mouse sperm. Sperm isolated from the cauda (+/+: wild-type, −/−: CRIS knockout) have been probed with IFT172- and KIF2A-specific antibodies and a fluorescent secondary antibody (green). DNA was stained with DAPI (blue). Scale bars are indicated.</p>", "links"=>[], "tags"=>["putative", "CRIS"], "article_id"=>870192, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.g007", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_putative_CRIS_interaction_partners_/870192", "title"=>"Expression of putative CRIS interaction partners.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305645"], "description"=>"<p>(<b>A</b>) Electron micrograph (cross section) of the midpiece (left) and principal piece (right) of the sperm flagellum from wild-type (+/+, top) and <i>Cris</i>-deficient (−/−, bottom) mice. (<b>B</b>) Minimal projection of a cross-section through the midpiece of the sperm flagellum from <i>Cris</i>-deficient (−/−) mice. The corresponding movie (Movie S1) can be found in the supplementary information. (<b>C</b>) Analysis of sperm motility. Velocity of straight line path (VSL) has been determined from the corresponding movies (Movie S2, S3). Data are presented as mean ± s.d. (<b>D</b>) Analysis of PKA- and tyrosine kinase-dependent protein phosphorylation under capacitating conditions and non-capacitating conditions. Per conditions, 1×10<sup>6</sup> cells were used. Immunoblots have been probed with PKA and tyrosine kinase substrate-specific (4G10) antibodies. Loading control: β-tubulin. (<b>E</b>) Ca<sup>2+</sup> imaging of mouse sperm loaded with caged 2-Diazo and Fluo-8. 2-Diazo was released using a UV flash. Shown is a representative trace of a wild-type sperm. Fluorescence has been background-subtracted and normalized to the value before the flash. (<b>F</b>) Flagellar waveform analysis of sperm from <i>Cris</i>-deficient mice loaded with caged 2-Diazo. Representative analysis showing the asymmetry index before and after the UV flash. Corresponding images of the flagellar waveform before and after the UV flash (yellow arrow) are included. (<b>G</b>) Mean values of the asymmetry index for wild-type (blue) and mutant sperm (black). Data are plotted as mean ± s.e.m. Corresponding movies (Movie S4, S5) can be found in the supplementary information (+/+: n = 24, −/−: n = 22). (<b>H</b>) Relative change of asymmetry index for wild-type (blue) and mutant sperm (black). Values after the UV flash were normalized to the values before the flash. Individual data are plotted as circles, the corresponding mean is represented by a red line, and the s.e.m. is indicated with a box, p = 0.0075.</p>", "links"=>[], "tags"=>["controls", "flagellar"], "article_id"=>870190, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.g006", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CRIS_controls_the_development_of_flagellar_bending_/870190", "title"=>"CRIS controls the development of flagellar bending.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305642"], "description"=>"<p>(<b>A</b>) Representative images showing the flagellar waveform of tethered non-capacitated wild-type (+/+) and mutant (−/−) sperm. Images were acquired at 200 frames per second. (<b>B</b>) Flagellar bending modes of mouse sperm. Left: pro-hook conformation, right: anti-hook conformation. (<b>C</b>) Quantification of the flagellar-waveform asymmetry for cells shown in panels A, D, and E. The height of the boxes indicates the s.d. around the mean (red). Big symbols on each boxplot are displayed in the same order (from left to right) as the cells shown on the corresponding panel. Small symbols represent cells that are not shown in the figure. Blue triangles pointing up (down) correspond to those cells on the top (bottom) of panel D. (<b>D</b>) Capacitated wild-type sperm alternate between bending in the pro- (bottom row) and anti-hook conformation (top row). (<b>E</b>) Capacitated mutant sperm remain in the anti-hook conformation.</p>", "links"=>[], "tags"=>[], "article_id"=>870187, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.g005", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Flagellar_waveform_/870187", "title"=>"Flagellar waveform.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305650"], "description"=>"<p>Phenotypic characterization of <i>Cris</i>-deficient mice.</p>", "links"=>[], "tags"=>["characterization"], "article_id"=>870195, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.t001", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phenotypic_characterization_of_Cris_deficient_mice_/870195", "title"=>"Phenotypic characterization of <i>Cris</i>-deficient mice.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305628"], "description"=>"<p>(<b>A</b>) Sequence comparison of CNBDs from different proteins. Sequence alignment of CNBDs from mCRIS, cyclic nucleotide-gated channels (bCNGA1, rCNGA4), a hyperpolarization activated and cyclic nucleotide-gated channel (mHCN2), a regulatory subunit from PKA (bPKARI-B), the exchange protein directly-activated by cAMP (hEPAC1), the bacterial catabolite activator protein (CAP), and the ELK1 channel from zebrafish (zELK). Amino acids that have been shown to be essential for ligand binding <a href=\"http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003960#pgen.1003960-Cukkemane1\" target=\"_blank\">[1]</a> are highlighted with asterisks. The β strand that functions as an intrinsic ligand in the ELK channels is highlighted in grey. Secondary structure elements are indicated below (β sheets: β 1–8, black arrows; α helices: αA–C, PBC, grey boxes). (<b>B</b>) M4T model of the presumed CNBD of mCRIS in the presence of cAMP. (<b>C</b>) Close-up view of the CNBD model of mCRIS indicating important interactions of side chain and backbone atoms with cAMP. (<b>D–G</b>) Analysis of cAMP binding using FRET. (<b>D</b>) Model demonstrating that binding of cAMP changes the conformation of the CNBD resulting in a change in FRET. (<b>E</b>) Representative traces for the change in cerulean (blue) and citrine (yellow) emission during perfusion of cit-mCNBD-cer expressing CHO cells with 3 mM 8-Br-cAMP. Arrow indicates start of perfusion. (<b>F</b>) Average change in FRET (normalized emission ratio cerulean/FRET-citrine) during perfusion of cit-mCNBD-cer expressing cells with 3 mM 8-Br-cAMP (CNBD 8-Br-cAMP), 40 µM NKH477/100 µM IBMX (CNBD NKH/IBMX), 3 mM 8-Br-cGMP (CNBD 8-Br-cGMP), and cit-mCNBD-R288Q-cer expressing cells with 3 mM 8-Br-cAMP (CNBD-RQ 8-Br-cAMP), and 40 µM NKH477/100 µM IBMX (CNBD-RQ NKH/IBMX). Arrow indicates start of perfusion. (<b>G</b>) Average change in FRET after 10 min of perfusion (mean ± s.d., black; 95% confident interval, dotted, grey). N numbers and p values are indicated.</p>", "links"=>[], "tags"=>[], "article_id"=>870177, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.g002", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CRIS_is_a_novel_target_for_cAMP_/870177", "title"=>"CRIS is a novel target for cAMP.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305649"], "description"=>"<p>Proteins identified by mass spectrometry.</p>", "links"=>[], "tags"=>[], "article_id"=>870194, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.t002", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Proteins_identified_by_mass_spectrometry_/870194", "title"=>"Proteins identified by mass spectrometry.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-12-05 04:52:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1305626"], "description"=>"<p>(<b>A</b>) Phylogenetic tree of CNBD-containing proteins. The different families have been labeled with different colors. Bootstrap values are shown as percentages. Scale bar shows amino acid substitution rate for the NJ (neighbor joining) tree. (<b>B</b>) Metazoan phylogeny describing the presence or absence of CRIS in metazoan genomes. The phylogenetic branching pattern was extracted from the Tree of Life project (<a href=\"http://www.toolweb.org/tree/\" target=\"_blank\">http://www.toolweb.org/tree/</a>) as of December 2012. The metazoan lineages known to contain CRIS are indicated by grey boxes, whereas those lineages that are believed to lack CRIS are indicated with white boxes.</p>", "links"=>[], "tags"=>["constitutes", "cnbd-containing"], "article_id"=>870175, "categories"=>["Biological Sciences"], "users"=>["Anke Miriam Krähling", "Luis Alvarez", "Katharina Debowski", "Qui Van", "Monika Gunkel", "Stephan Irsen", "Ashraf Al-Amoudi", "Timo Strünker", "Elisabeth Kremmer", "Eberhard Krause", "Ingo Voigt", "Simone Wörtge", "Ari Waisman", "Ingo Weyand", "Reinhard Seifert", "Ulrich Benjamin Kaupp", "Dagmar Wachten"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1003960.g001", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CRIS_constitutes_a_new_member_of_the_CNBD_containing_protein_family_/870175", "title"=>"CRIS constitutes a new member of the CNBD-containing protein family.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-05 04:52:22"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences/Biomechanics", "average_usage"=>[262, 443, 579, 690, 795, 884, 991, 1089, 1183, 1275, 1366, 1445, 1522]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[272, 472, 600, 713, 815, 911, 1004, 1094, 1185, 1273, 1358, 1441]}, {"subject_area"=>"/Biology and life sciences/Physiology", "average_usage"=>[256, 449, 572, 676, 775, 866, 955, 1041, 1127, 1213, 1290, 1370, 1437]}, {"subject_area"=>"/Medicine and health sciences", "average_usage"=>[264, 460, 584, 692, 794, 887, 978, 1067, 1154, 1241, 1328, 1408, 1474]}, {"subject_area"=>"/Medicine and health sciences/Pathology and laboratory medicine", "average_usage"=>[267, 466, 592, 709, 806, 901, 989, 1075, 1162, 1254, 1342, 1424, 1486]}, {"subject_area"=>"/Medicine and health sciences/Urology", "average_usage"=>[264, 490, 621, 739, 833, 927, 1020, 1102, 1204, 1282, 1349, 1432, 1504]}, {"subject_area"=>"/Physical sciences/Physics", "average_usage"=>[254, 421, 527, 626, 720, 813, 900, 983, 1063, 1136, 1210, 1283, 1342]}]}
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Net::HTTPTooManyRequests

Source
Scopus
Time
2019-08-27 19:05:11 UTC
Target URL
https://api.elsevier.com/content/search/index:SCOPUS?query=DOI(10.1371%2Fjournal.pgen.1003960)
Trace

/app/models/concerns/networkable.rb:21:in `get_result'
/app/models/source.rb:165:in `get_data'
/app/models/retrieval_status.rb:47:in `perform_get_data'
/app/jobs/source_job.rb:52:in `block (2 levels) in perform'
/app/jobs/source_job.rb:51:in `block in perform'
/app/jobs/source_job.rb:35:in `each'
/app/jobs/source_job.rb:35:in `perform'