Targeting and Anchoring Tudor in the Pole Plasm of the Drosophila Oocyte
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{"title"=>"Targeting and anchoring Tudor in the pole plasm of the Drosophila oocyte", "type"=>"journal", "authors"=>[{"first_name"=>"Joël", "last_name"=>"Anne", "scopus_author_id"=>"56256010300"}], "year"=>2010, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"21179512", "doi"=>"10.1371/journal.pone.0014362", "sgr"=>"78650721217", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "scopus"=>"2-s2.0-78650721217", "issn"=>"19326203", "pui"=>"361019998"}, "id"=>"53e97c5f-6bcc-362f-a8ac-3f9ca6c343d9", "abstract"=>"Germline formation is a highly regulated process in all organisms. In Drosophila embryos germ cells are specified by the pole plasm, a specialized cytoplasmic region containing polar granules. Components of these granules are also present in the perinuclear ring surrounding nurse cells, the nuage. Two such molecules are the Vasa and Tudor proteins. How Tudor localizes and is maintained in the pole plasm is, however, not known.", "link"=>"http://www.mendeley.com/research/targeting-anchoring-tudor-pole-plasm-drosophila-oocyte", "reader_count"=>32, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>2, "Student > Master"=>5, "Other"=>1, "Student > Bachelor"=>3, "Lecturer > Senior Lecturer"=>2}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>2, "Student > Master"=>5, "Other"=>1, "Student > Bachelor"=>3, "Lecturer > Senior Lecturer"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>6, "Agricultural and Biological Sciences"=>25}, "reader_count_by_subdiscipline"=>{"Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>25}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"United States"=>1, "Germany"=>1, "Spain"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/813279"], "description"=>"<p>(A-C) Distribution of GFP-Vas (green) and Tud (red) in wild-type (A) left, stage 6, and right, stage 8, (B) stage 9, and (C) stage 10 egg chambers. (A, upper panel) In stage 6 egg chambers Tud was enriched in the oocyte and in stage 8 transiently accumulated at the anterior margin of the oocyte, whereas GFP-Vas was predominantly detected in the nurse cells. (Lower panel) Both GFP-Vas and Tud decorated the membrane of nurse cell nuclei but the distribution of both proteins did not fully overlap; some Vas-containing particles were free of Tud. (B). In stage 9 egg chambers Tud and GFP-Vas began to accumulate at the posterior pole of the oocyte. (C, upper panel) Localization of GFP-Vas and Tud in nuage and pole plasm overlapped in a stage 10 egg chamber. (Lower panel) Higher magnification of the oocyte posterior cortex reveals the occurrence of particles containing both GFP-Vas and Tud in close proximity to the pole plasm (arrows). (D) RNA-dependent association of Tud with Vas. Immuno-detection of Tud in ovarian protein extracts separated by SDS-PAGE and blotted on PVDF membrane of wild-type females (first lane), homozygous <i>tud<sup>1</sup></i> females (second lane), and affinity purified Vas-complexes isolated in presence (third lane) or absence of RNase inhibitors (fourth lane). Vas-complexes were purified from ovarian extracts representing ∼25-fold the amount of proteins loaded in the first lane. (E) RNase sensitive binding of Tud to Vas-complexes. Affinity purified Vas-complexes were isolated from ovarian protein extracts representing ∼75 fold the amount used in the first lane, as control, in presence of RNase inhibitors. Following purification the Vas-complexes were treated with RNase A and the Vas-complexes were isolated by centrifugation. Proteins in the pellet (P) and the supernatant (S) were separated and similarly analyzed for the occurrence of Tud as in D.</p>", "links"=>[], "tags"=>["tud", "vas"], "article_id"=>483656, "categories"=>["Developmental Biology"], "users"=>["Joël Anne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014362.g005", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Relationship_between_Tud_and_Vas_during_oogenesis_/483656", "title"=>"Relationship between Tud and Vas during oogenesis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-12-15 01:00:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/813094"], "description"=>"<p>Distribution of Tud in (A) wild-type and (B) <i>osk<sup>84</sup></i>/<i>Df(3R)pXT103</i> stage 10 egg chambers. In an <i>osk</i>-protein null egg chamber Tud localization in the pole plasm was abolished.</p>", "links"=>[], "tags"=>["tud", "osk"], "article_id"=>483465, "categories"=>["Developmental Biology"], "users"=>["Joël Anne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014362.g003", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Recruitment_of_Tud_by_Osk_protein_/483465", "title"=>"Recruitment of Tud by Osk protein.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-12-15 00:57:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/812960"], "description"=>"<p>Distribution of Tud in (A) wild-type, (B) <i>vas<sup>Q7</sup></i>, (C) <i>vas<sup>O11</sup></i>, (D) <i>aub<sup>HN/N11</sup></i>, (E) <i>krimp<sup>Qf065837</sup></i>/<i>Df (2R)Exel<sup>6063</sup>,</i> and (F) <i>mael<sup>M391</sup></i>/<i>Df(3L)79E-F</i> stage 10 egg chambers and early cleavage embryos. Tud (red), DNA (green). Higher magnifications of the posterior cortex of the oocyte are provided on the right panels. (A, upper panel) In wild-type Tud forms a crescent at the posterior of the oocyte in stage 10 egg chamber and (lower panel) is maintained at the posterior pole during early embryogenesis. (B) In the strong <i>vas<sup>Q7</sup></i> allele Tud accumulated at the posterior pole but became partially dissociated from the oocyte cortex and frequently formed a ring structure around hypothetical yolk particles. This structure stained negatively for lamin and thus did not correspond to the oocyte nucleus (data not shown). (C, upper panel) In the weaker <i>vas<sup>O11</sup></i> allele Tud was poorly associated with the cortex and (lower panel) absent from the pole plasm during early embryogenesis. In (D) <i>aub<sup>HN/N11</sup></i> and (E) <i>krimp<sup>Qf065837</sup></i>/<i>Df(2R)Exel<sup>6063</sup></i> stage 10 egg chambers Tud was associated with particles dispersed in the bulk cytoplasm. A fraction of the Tud particles formed a loose association with the cortex at the posterior pole. (F) In a <i>mael<sup>M391</sup></i>/<i>Df(3L)79E-F</i> stage 10 egg chamber a weak crescent of Tud was associated with the posterior pole cortex through pillar structures.</p>", "links"=>[], "tags"=>["nuage", "tud"], "article_id"=>483338, "categories"=>["Developmental Biology"], "users"=>["Joël Anne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014362.g002", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Components_of_the_nuage_are_required_for_the_stability_of_Tud_in_the_pole_plasm_/483338", "title"=>"Components of the nuage are required for the stability of Tud in the pole plasm.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-12-15 00:55:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/812842"], "description"=>"<p>Distribution of Tud and DNA in (A) wild-type, (B) <i>vas<sup>Q7</sup></i>, (C) <i>vas<sup>O11</sup></i>, (D) <i>aub<sup>HN/N11</sup></i>, (E), <i>mael<sup>M391</sup></i>/<i>Df(3L)79E-F</i>, and (F) <i>krimp<sup>Qf065837</sup></i>/<i>Df(2R)Exel<sup>6063</sup></i> stage 8 egg chambers. Tud (red), DNA (green). The right column displays a higher magnification of Tud distribution around wild-type and mutant nurse cell nuclei. (A) Tud normally accumulates at the periphery of the nurse cell nucleus. (B) Tud is absent from the nuage in the strong <i>vas<sup>Q7</sup></i> allele and (C) is reduced in the weaker <i>vas<sup>O11</sup></i> allele. (D) In <i>aub<sup>HN/N11</sup></i> Tud aggregates in a single large spot bound to the nuclear membrane. (E) In <i>mael<sup>M391</sup></i>/<i>Df(3L)79E-F</i> Tud accumulation in the nuage is severely impaired (left panel) but small aggregates containing Tud could be detected in the cytoplasm close to the nuclear membrane (right panel). (F) Tud localization in the vicinity of the nuclear membrane is drastically reduced in <i>krimp<sup>Qf065837</sup></i>/<i>Df(2R)Exel<sup>6063</sup></i> egg chambers.</p>", "links"=>[], "tags"=>["tud", "localization"], "article_id"=>483214, "categories"=>["Developmental Biology"], "users"=>["Joël Anne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014362.g001", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Genetic_requirement_for_Tud_localization_in_the_nuage_/483214", "title"=>"Genetic requirement for Tud localization in the nuage.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-12-15 00:53:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/813387"], "description"=>"<p>Full length GST-Vas (648 amino acid residues) or derivatives were purified from bacterial extracts and incubated with S•Tag-Osk. (Left, upper panels) Following separation by SDS-PAGE the bound S•Tag-Osk proteins were detected by immuno-blotting using alkaline phosphatase-conjugated S proteins. Input: one tenth of protein extract was loaded on the gel. (Lower panel) The amount of GST-Vas proteins was visualized by Coomassie staining. (Right) Representation of the GST-Vas fragments used for mapping and summary of the results. The RG repeats are indicated by black circles and the Gustavus- and Osk-binding domains of Vas are depicted as yellow and red boxes, respectively.</p>", "links"=>[], "tags"=>["osk-binding"], "article_id"=>483765, "categories"=>["Developmental Biology"], "users"=>["Joël Anne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014362.g006", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Delimitation_of_the_Osk_binding_domain_in_Vas_/483765", "title"=>"Delimitation of the Osk-binding domain in Vas.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-12-15 01:02:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/813460"], "description"=>"<p>This model is mainly based on the finding that Tud and Vas can localize in polar granule independently from each other and that Vls mediates the localization of Tud by binding to both Tud and Osk. An association between Vas and Tud could be mediated by RNA or through protein-protein interaction (See <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0014362#s3\" target=\"_blank\">Discussion</a>).</p>", "links"=>[], "tags"=>["polar", "granule"], "article_id"=>483833, "categories"=>["Developmental Biology"], "users"=>["Joël Anne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014362.g007", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Scheme_of_polar_granule_assembly_/483833", "title"=>"Scheme of polar granule assembly.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-12-15 01:03:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/405257", "https://ndownloader.figshare.com/files/405264", "https://ndownloader.figshare.com/files/405275"], "description"=>"<div><h3>Background</h3><p>Germline formation is a highly regulated process in all organisms. In <em>Drosophila</em> embryos germ cells are specified by the pole plasm, a specialized cytoplasmic region containing polar granules. Components of these granules are also present in the perinuclear ring surrounding nurse cells, the nuage. Two such molecules are the Vasa and Tudor proteins. How Tudor localizes and is maintained in the pole plasm is, however, not known.</p><h3>Methodology/Principal Findings</h3><p>Here, the process of Tudor localization in nuage and pole plasm was analyzed. The initial positioning of Tudor at the posterior pole of stage 9 oocytes was found to occur in the absence of a structurally detectable nuage. However, in mutants for genes encoding components of the nuage, including <em>vasa, aubergine, maelstrom,</em> and <em>krimper,</em> Tudor was detached from the posterior cortex in stage 10 oocytes, suggesting a prior passage in the nuage for its stability in the pole plasm. Further studies indicated that Valois, which was previously shown to bind <em>in vitro</em> to Tudor, mediates the localization of Tudor in the pole plasm by physically interacting with Oskar, the polar granule organizer. An association between Tudor and Vasa mediated by RNA was also detected in ovarian extracts.</p><h3>Conclusions/Significance</h3><p>The present data challenge the view that the assembly of the polar granules occurs in a stepwise and hierarchical manner and, consequently, a revised model of polar granule assembly is proposed. In this model Oskar recruits two downstream components of the polar granules, Vasa and Tudor, independently from each other: Vasa directly interacts with Oskar while Valois mediates the recruitment of Tudor by interacting with Oskar and Tudor.</p></div>", "links"=>[], "tags"=>["targeting", "anchoring", "tudor", "plasm", "oocyte"], "article_id"=>140089, "categories"=>["Developmental Biology"], "users"=>["Joël Anne"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0014362.s001", "https://dx.doi.org/10.1371/journal.pone.0014362.s002", "https://dx.doi.org/10.1371/journal.pone.0014362.s003"], "stats"=>{"downloads"=>5, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Targeting_and_Anchoring_Tudor_in_the_Pole_Plasm_of_the_Drosophila_Oocyte/140089", "title"=>"Targeting and Anchoring Tudor in the Pole Plasm of the <em>Drosophila</em> Oocyte", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2010-12-15 00:01:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/813174"], "description"=>"<p>(A) Osk-binding domains in Vls. (Left, upper panel) Full length GST-Vls (367 amino acid residues) or derivatives were purified from bacterial extracts and the relative amounts of GST-fusion proteins were evaluated by SDS-PAGE followed by Coomassie staining. Amino acid numbers are given across the top. (Middle panel) The GST-fusion proteins were incubated with S•Tag-Osk. Bound S•Tag-Osk proteins were separated by SDS-PAGE electrophoresis and detected by immuno-blotting using alkaline phosphatase-conjugated S proteins. Input: one tenth of the protein extract was loaded on the gel. (Lower panel) To define more precisely the Osk-binding domain at the C-terminus of Vls, synthetic peptides corresponding to amino acid 310–340 and 340–367 of Vls were added to the binding assay containing the GST-Vls<sup>309-367</sup> fusion fragment. The peptide 310-340 inhibited Osk binding to GST-Vls<sup>309-367</sup>. (Right) Representation of the GST-Vls fragments used for the mapping and summary of the results. The four WD-repeats are numbered and the putative Osk-binding domains of Vls are depicted in green. (B) Two internal regions in Osk are necessary for its binding to Vls. Full size S•Tag-short Osk isoform (447 amino acid residues) or derivatives were synthesized in vitro and incubated with full-size GST-Vls. Following separation by SDS-PAGE electrophoresis the bound S•Tag-Osk proteins were detected by immuno-blotting using alkaline phosphatase-conjugated S proteins. (Left panel) Input S•Tag-Osk proteins. (Middle panel) Bound S•Tag-Osk proteins. (Right) Representation of S•Tag-Osk constructs used for the mapping and summary of the results with the identified domains required for Vls-binding in Osk shown in pink.</p>", "links"=>[], "tags"=>["interacts", "osk"], "article_id"=>483547, "categories"=>["Developmental Biology"], "users"=>["Joël Anne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014362.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Vls_interacts_physically_with_the_short_Osk_isoform_/483547", "title"=>"Vls interacts physically with the short Osk isoform.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-12-15 00:59:07"}

PMC Usage Stats | Further Information

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