The Role of 14-3-3ε Interaction with Phosphorylated Cdc25B at Its Ser321 in the Release of the Mouse Oocyte from Prophase I Arrest
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{"title"=>"The Role of 14-3-3ε Interaction with Phosphorylated Cdc25B at Its Ser321 in the Release of the Mouse Oocyte from Prophase I Arrest", "type"=>"journal", "authors"=>[{"first_name"=>"Jun", "last_name"=>"Meng", "scopus_author_id"=>"56325925100"}, {"first_name"=>"Cheng", "last_name"=>"Cui", "scopus_author_id"=>"22955273400"}, {"first_name"=>"Yanchun", "last_name"=>"Liu", "scopus_author_id"=>"55554934100"}, {"first_name"=>"Minglin", "last_name"=>"Jin", "scopus_author_id"=>"55554472600"}, {"first_name"=>"Didi", "last_name"=>"Wu", "scopus_author_id"=>"55554185000"}, {"first_name"=>"Chao", "last_name"=>"Liu", "scopus_author_id"=>"56938146100"}, {"first_name"=>"Enhua", "last_name"=>"Wang", "scopus_author_id"=>"7403414327"}, {"first_name"=>"Bingzhi", "last_name"=>"Yu", "scopus_author_id"=>"7402092717"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)", "pui"=>"368101302", "sgr"=>"84872255347", "doi"=>"10.1371/journal.pone.0053633", "scopus"=>"2-s2.0-84872255347", "pmid"=>"23326474"}, "id"=>"7bc655ee-84c2-354b-9c04-2649b310b8b2", "abstract"=>"The protein kinase A (PKA)/Cdc25B pathway plays a critical role in maintaining meiotic arrest in mouse oocytes. However, the molecular mechanism underlying this interchange is not known. In this study, we assessed the role of 14-3-3ε interaction with phosphorylated Cdc25B at its Ser321 as the mouse oocyte is released from prophase I arrest. The 14-3-3ε isoform is a highly conserved protein with various regulatory roles, including maintenance of meiotic arrest. Cdc25B phosphatase is also a key cell cycle regulator. 14-3-3ε binds to Cdc25B-WT, which was abrogated when Ser321 of Cdc25B was mutated to Ala. In addition, we found that 14-3-3ε and Cdc25B were co-localized. Cdc25B was translocated from the cytoplasm to the nucleus shortly before germinal vesicle breakdown (GVBD) during the primary oocyte stage of oogenesis. However, mutation of Ser321 to Ala completely abolished the cytoplasmic localization of Cdc25B. Furthermore, oocytes co-expressing of Cdc25B-WT or Cdc25B-Ser321D and 14-3-3ε were unable to undergo GVBD. In contrast, co-expression of 14-3-3ε and Cdc25B-Ser321A induced GVBD and allowed the process to continue. Down-regulation of 14-3-3ε caused partial meiotic resumption. Taken together, these data indicate that Ser321 of Cdc25B is the specific binding site for 14-3-3ε binding, and that 14-3-3ε is the significant factor in Cdc25B regulation during meiotic resumption of GV stage.", "link"=>"http://www.mendeley.com/research/role-1433%CE%B5-interaction-phosphorylated-cdc25b-ser321-release-mouse-oocyte-prophase-i-arrest", "reader_count"=>14, "reader_count_by_academic_status"=>{"Researcher"=>1, "Student > Ph. D. Student"=>5, "Student > Master"=>3, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_user_role"=>{"Researcher"=>1, "Student > Ph. D. Student"=>5, "Student > Master"=>3, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>7, "Agricultural and Biological Sciences"=>4, "Medicine and Dentistry"=>2, "Psychology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Psychology"=>{"Psychology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>4}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>7}}, "reader_count_by_country"=>{"South Korea"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/509821"], "description"=>"<p>(A) the mRNA levels of 14-3-3 isoforms at GV stage of mouse oocytes. mRNA of 120 mouse oocytes of GV stage were extracted (described in the“<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053633#s4\" target=\"_blank\">Materials and Methods</a>” Section). RT-PCR products using primers for specific 14-3-3 isoforms are observed in ethidium bromide-stained agarose gel. ε, β, γ, η, σ, τ and ζ represent seven 14-3-3 isoforms. (B) the mRNA levels of 14-3-3ε at GV and GVBD stage of mouse oocytes. Line GV: mouse oocytes at GV stage. Line GVBD: mouse oocytes at GVBD stage. (C) A protein band of approximately 28 kDa from 200 mouse oocytes at GV or GVBD stage is detectable by Western blot with an anti-14-3-3ε antibody (upper panel). In contrast an anti-β-actin antibody detected a low molecular weight band of approximately 43 kDa (lower panel). (D) Western blot analysis 14-3-3β protein expression with an anti-14-3-3β antibody from 300 mouse oocytes at GV or GVBD stage, A protein band of approximately 29 kDa (upper panel). Line Brain: mouse brain protein as positive control. Shown is a representative of three independent experiments.</p>", "links"=>[], "tags"=>["14-3-3", "isoforms", "gv", "gvbd"], "article_id"=>180304, "categories"=>["Physiology", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Jun Meng", "Cheng Cui", "Yanchun Liu", "Minglin Jin", "Didi Wu", "Chao Liu", "Enhua Wang", "Bingzhi Yu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0053633.g001", "stats"=>{"downloads"=>0, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Identification_of_14_3_3_isoforms_expressed_in_GV_and_GVBD_mouse_oocytes_/180304", "title"=>"Identification of 14-3-3 isoforms expressed in GV and GVBD mouse oocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 17:16:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/510127"], "description"=>"<p>(A) red fluorescent Cdc25B signals and green fluorescent 14-3-3ε signals were colocalized in the cytoplasm in GV-stage oocytes. (B) the partial red fluorescent signals were translocated to the nucleus in oocytes before GVBD (C) after GVBD, the red fluorescent signals and the green fluorescent signals were evenly distributed in the whole cell. Scale bar = 20 µm.</p>", "links"=>[], "tags"=>["endogenous", "cdc25b"], "article_id"=>180613, "categories"=>["Physiology", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Jun Meng", "Cheng Cui", "Yanchun Liu", "Minglin Jin", "Didi Wu", "Chao Liu", "Enhua Wang", "Bingzhi Yu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0053633.g004", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Co_localization_of_endogenous_Cdc25B_and_14_3_3_949_/180613", "title"=>"Co-localization of endogenous Cdc25B and 14-3-3ε.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 17:18:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/510031"], "description"=>"<p>Effects of microinjection with HA-tagged-14-3-3ε mRNA solely or co-injection with MYC-tagged-Cdc25B-WT mRNA, MYC-tagged-Cdc25B-Ser321A mRNA or MYC-tagged-Cdc25B-Ser321D mRNA and HA-tagged- 14-3-3ε mRNA on meiotic resumption of mouse oocytes. Experiments were performed in the continued presence of dbcAMP(A–G). HEK293 cells were transfected with 14-3-3ε together with empty vector, Cdc25B-WT , Cdc25B-Ser321A or Cdc25B-Ser321D (H). (A) at the indicated times, the percentages of germinal vesicle breakdown (GVBD) or MII or death were counted in cultured mouse oocytes after various mRNAs microinjection. GVBD, 3 h; MII or death, 20 h. The total number of oocytes undergoing meiotic maturation or death is given on top of bar graph from three independent experiments. (B) the GVBD rate at indicated time points in cultured mouse oocytes after various mRNAs microinjection. (C) H1 kinase activity in oocytes injected with various mRNAs. Each value was expressed as mean±S.D of at least thee independent experiments. (D) Protein extracts from oocytes microinjected with various mRNAs were incubated with histone H1 and [γ<sup>−32</sup>P] ATP. The protein was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and incorporation of <sup>32</sup>P into histone H1 was visualized by autoradiography. (E) Western analysis of phosphorylation status of Cdc2-Tyr15. GV oocytes injected with various mRNAs were collected 2 h after microinjection. The collected oocytes were immunoblotted with anti-pTyr15 of Cdc2 antibody. (F) Western analysis of Cdc25B-MYC and endogenous Cdc25B expression. The oocytes co-injected with Cdc25B-WT mRNA, Cdc25B-Ser321A mRNA or Cdc25B-Ser321D mRNA and 14-3-3ε mRNA were collected 3 h after injection and their proteins were immunoblotted with anti-myc, anti-Cdc25B or anti-beta-actin antibody. Bars represent means±S.D of thee independent experiments. (G) Western analysis of HA-14-3-3ε and endogenous 14-3-3ε expression. The oocytes injected with 14-3-3ε mRNA solely or co-injected with Cdc25B-WT mRNA, Cdc25B-Ser321A mRNA or Cdc25B-Ser321D mRNA and 14-3-3ε mRNA were collected 3 h after injection and their proteins were immunoblotted with anti-HA, anti-14-3-3ε or anti-beta-actin antibody. Bars represent means±S.D of thee independent experiments. (H) Western blot analysis of Cdc25B interaction with 14-3-3ε (IP) and Western blot analysis demonstrating the expression of HA-14-3-3ε and EGFP-Cdc25B in the cell lysates used to immunoprecipitate Cdc25B protein shown in (IP), an anti-GAPDH antibody was used to determine equal loading of the gel.</p>", "links"=>[], "tags"=>["cdc25b"], "article_id"=>180524, "categories"=>["Physiology", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Jun Meng", "Cheng Cui", "Yanchun Liu", "Minglin Jin", "Didi Wu", "Chao Liu", "Enhua Wang", "Bingzhi Yu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0053633.g003", "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ser321_of_Cdc25B_is_the_sole_site_responsible_for_14_3_3_949_binding_/180524", "title"=>"Ser321 of Cdc25B is the sole site responsible for 14-3-3ε binding.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 17:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/510212"], "description"=>"<p>(A) red fluorescent 14-3-3ε signals and green fluorescent Cdc25B-WT signals were co-localized in the cytoplasm in GV-stage oocytes. (B) red fluorescent 14-3-3ε signals and green fluorescent Cdc25B-Ser321A signals were distributed in the whole cell, but particularly dense in the nucleus. Scale bar = 20 µm.</p>", "links"=>[], "tags"=>["exogenously", "cdc25b"], "article_id"=>180709, "categories"=>["Physiology", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Jun Meng", "Cheng Cui", "Yanchun Liu", "Minglin Jin", "Didi Wu", "Chao Liu", "Enhua Wang", "Bingzhi Yu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0053633.g005", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Co_localization_of_exogenously_expressed_Cdc25B_and_14_3_3_949_/180709", "title"=>"Co-localization of exogenously expressed Cdc25B and 14-3-3ε.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 17:18:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/509929"], "description"=>"<p>(A) 120 GV-oocytes microinjected with 14-3-3ε siRNA or control siRNA (5pl of 20 µM) were collected 24 h after microinjection. Oocytes microinjected with control siRNA and no injection served as control. mRNA of 14-3-3ε was detected by RT-PCR using primers specific for 14-3-3ε. (B) Protein expression of 14-3-3ε was examined by western blot with an anti-14-3-3ε antibody. β-actin used as endogenous internal reference demonstrate 14-3-3ε siRNA specificity and equal loading. (C) at the indicated times, the percentages of germinal vesicle breakdown (GVBD) or MII or death were counted in cultured mouse oocytes after 14-3-3ε siRNA or control siRNA microinjection. GVBD, 24 h; MII or death, 30 h. The total number of oocytes undergoing meiotic maturation or death is given on top of bar graph from three independent experiments. (D) H1 kinase activity in oocytes injected with 14-3-3ε siRNA or control siRNA or no injection groups. Each value was expressed as mean±S.D of at least three independent experiments. (E) Protein extracts from oocytes microinjected with 14-3-3ε siRNA or control siRNA and no injection groups were incubated with histone H1 and [γ<sup>-32</sup>P] ATP. The protein was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and incorporation of <sup>32</sup>P into histone H1 was visualized by autoradiography. (F) Western analysis of phosphorylation status of Cdc2-Tyr15. GV oocytes injected with 14-3-3ε siRNA or control siRNA were collected 24 h after microinjection. The collected oocytes were immunoblotted with anti-pTyr15 of Cdc2 antibody. (G) Co-localization of Endogenous Cdc25B and 14-3-3ε in mouse oocytes injected with 14-3-3ε siRNA or control siRNA. red fluorescent Cdc25B signals and green fluorescent 14-3-3ε signals were co-localized in the cytoplasm in oocytes injected with control siRNA, while red fluorescent Cdc25B signals were translocated from the cytoplasm to the nucleus in oocytes injected with14-3-3ε siRNA. Scale bar = 20 µm.</p>", "links"=>[], "tags"=>["sirna", "causes", "meiotic", "resumption"], "article_id"=>180415, "categories"=>["Physiology", "Cell Biology", "Genetics", "Developmental Biology"], "users"=>["Jun Meng", "Cheng Cui", "Yanchun Liu", "Minglin Jin", "Didi Wu", "Chao Liu", "Enhua Wang", "Bingzhi Yu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0053633.g002", "stats"=>{"downloads"=>9, "page_views"=>31, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Knockdown_of_14_3_3_949_by_siRNA_causes_partial_meiotic_resumption_mouse_oocytes_/180415", "title"=>"Knockdown of 14-3-3ε by siRNA causes partial meiotic resumption mouse oocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 17:17:09"}

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", "average_usage"=>[269, 466, 588, 697, 800, 896, 988, 1076, 1165, 1254, 1340, 1417]}, {"subject_area"=>"/Biology and life sciences/Biochemistry", "average_usage"=>[266, 468, 593, 703, 804, 903, 993, 1084, 1171, 1256, 1339, 1422, 1492]}, {"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/Genetics", "average_usage"=>[284, 491, 620, 738, 843, 945, 1043, 1137, 1225, 1315, 1400, 1479, 1555]}, {"subject_area"=>"/Biology and life sciences/Organisms", "average_usage"=>[281, 484, 611, 728, 835, 934, 1030, 1123, 1214, 1299, 1383, 1464]}]}
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