Evidence for OTUD-6B Participation in B Lymphocytes Cell Cycle after Cytokine Stimulation
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{"title"=>"Evidence for OTUD-6B participation in B lymphocytes cell cycle after cytokine stimulation", "type"=>"journal", "authors"=>[{"first_name"=>"Zhongping", "last_name"=>"Xu", "scopus_author_id"=>"7405428196"}, {"first_name"=>"Yufang", "last_name"=>"Zheng", "scopus_author_id"=>"7404837356"}, {"first_name"=>"Yufei", "last_name"=>"Zhu", "scopus_author_id"=>"56052222700"}, {"first_name"=>"Xiangyin", "last_name"=>"Kong", "scopus_author_id"=>"15742790800"}, {"first_name"=>"Landian", "last_name"=>"Hu", "scopus_author_id"=>"7401556896"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-79251537162", "pmid"=>"21267069", "issn"=>"19326203", "sgr"=>"79251537162", "doi"=>"10.1371/journal.pone.0014514", "pui"=>"361185822"}, "id"=>"931f16fe-46bc-3176-a623-62cfd3ec40b9", "abstract"=>"Deubiquitinating enzymes (DUBs) are important regulators of cell proliferation. Here we identified a functional deubiquitinating enzyme, ovarian tumor domain-containing 6B (OTUD-6B). Mutation of the conserved Cys residue abolished its deubiquitinating activity in vitro. Otud-6b expression was induced with cytokine stimulation in both mouse Ba/F3 cells and primary B lymphocytes followed a rapid decrease. This rapid decrease was partially facilitated by tristetraprolin (TTP) destabilization of Otud-6b mRNA through AU-rich motifs. Enforced expression of OTUD-6B in Ba/F3 cells could block cell proliferation by arresting cells in G1 phase. In addition, cyclin D2 level was down-regulated when OTUD-6B WT was overexpressed. Therefore, down-regulation of Otud-6b expression after prolonged cytokine stimulation may be required for cell proliferation in B lymphocytes.", "link"=>"http://www.mendeley.com/research/evidence-otud6b-participation-b-lymphocytes-cell-cycle-after-cytokine-stimulation", "reader_count"=>6, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>2, "Student > Master"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>2, "Student > Master"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Agricultural and Biological Sciences"=>2, "Physics and Astronomy"=>1, "Chemistry"=>2}, "reader_count_by_subdiscipline"=>{"Chemistry"=>{"Chemistry"=>2}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"United States"=>1, "United Kingdom"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/805819"], "description"=>"<p><b>A.</b> The sequence of Otud-6b 3′UTR. AUUUA pentamer or AUUUUA hexamer are underlined. Each rectangle indicates an AU-rich sequence site. The AU-rich sequence with U surrounding it is indicated as a black rectangle. <b>B.</b> pSUPER Mock, pSUPER siTTP-1 or 2 and HA-TTP vectors co-expressing Ba/F3 cells were seeded at 2×10<sup>5</sup> cells per ml. Cell extracts were immunoblotted with anti-HA antibody. GFP was used as a loading control. <b>C.</b> Otud-6b RNA level in Ba/F3 cells transfected with pSUPER siTTP-1,2 vectors after starvation and stimulation with 10 pM mouse IL-3 for the indicated times (0, 0.5, 1, 2, 4, 8, 12, 16, and 24 hours). <b>D.</b> Ba/F3 cells were transfected with pSUPER Mock, pSUPER siTTP-1 or 2 vectors. 24 later, cells were stimulated by 10 pM IL-3 and incubated with 15 ug/ml Act.D for the time indicated. The qRT-PCR was performed to detect Otud-6b mRNA remaining at each time point. <b>E.</b> Semi-quantitative RT-PCR analysis of Otud-6b mRNA remaining at each time point after Act.D adding. <b>F.</b> RT-PCR for Otud-6b mRNA in protein-mRNA immunoprecipitation complex. The mRNAs of Ba/F3 cells transfected with HA-TTP when response to IL-3 was immunoprecipitated with anti-HA and IgG serum. As a control, Input (10%) denoted that the RNA sample contained Otud-6b mRNA and didn't contain genomic DNA. <b>G.</b> TTP transfection had little effect on pGL3 control luciferase expression. Wild-type TTP transfection resulted in a significant decrease in full-length Otud-6b 3′-UTR luciferase reporter expression in 293T cells, while the TTP Zn-FM did not alter Otud-6b 3′-UTR-mediated expression. Different region of Otud-6b 3′UTR was inserted into downstream of luciferase coding region. The truncate 3′UTR vectors were constructed depending on the AU-rich sequence distribution. 293T cells were provided by the Cell Bank, Shanghai Institute of Biochemistry and Cell Biology, SIBS, CAS.</p>", "links"=>[], "tags"=>["otud-6b", "mrna", "degradation"], "article_id"=>476190, "categories"=>["Cell Biology"], "users"=>["Zhongping Xu", "Yufang Zheng", "Yufei Zhu", "Xiangyin Kong", "Landian Hu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014514.g005", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TTP_is_involved_in_Otud_6b_mRNA_rapid_degradation_after_induction_/476190", "title"=>"TTP is involved in Otud-6b mRNA rapid degradation after induction.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-01-18 01:43:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/402376", "https://ndownloader.figshare.com/files/402413", "https://ndownloader.figshare.com/files/402426", "https://ndownloader.figshare.com/files/402442", "https://ndownloader.figshare.com/files/402458", "https://ndownloader.figshare.com/files/402480", "https://ndownloader.figshare.com/files/402501", "https://ndownloader.figshare.com/files/402523", "https://ndownloader.figshare.com/files/402545", "https://ndownloader.figshare.com/files/402563", "https://ndownloader.figshare.com/files/402577", "https://ndownloader.figshare.com/files/402596", "https://ndownloader.figshare.com/files/402626", "https://ndownloader.figshare.com/files/402659"], "description"=>"<div><p>Deubiquitinating enzymes (DUBs) are important regulators of cell proliferation. Here we identified a functional deubiquitinating enzyme, ovarian tumor domain-containing 6B (OTUD-6B). Mutation of the conserved Cys residue abolished its deubiquitinating activity <em>in vitro</em>. Otud-6b expression was induced with cytokine stimulation in both mouse Ba/F3 cells and primary B lymphocytes followed a rapid decrease. This rapid decrease was partially facilitated by tristetraprolin (TTP) destabilization of Otud-6b mRNA through AU-rich motifs. Enforced expression of OTUD-6B in Ba/F3 cells could block cell proliferation by arresting cells in G1 phase. In addition, cyclin D2 level was down-regulated when OTUD-6B WT was overexpressed. Therefore, down-regulation of Otud-6b expression after prolonged cytokine stimulation may be required for cell proliferation in B lymphocytes.</p></div>", "links"=>[], "tags"=>["otud-6b", "lymphocytes", "cytokine", "stimulation"], "article_id"=>139515, "categories"=>["Cell Biology"], "users"=>["Zhongping Xu", "Yufang Zheng", "Yufei Zhu", "Xiangyin Kong", "Landian Hu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0014514.s001", "https://dx.doi.org/10.1371/journal.pone.0014514.s002", "https://dx.doi.org/10.1371/journal.pone.0014514.s003", "https://dx.doi.org/10.1371/journal.pone.0014514.s004", "https://dx.doi.org/10.1371/journal.pone.0014514.s005", "https://dx.doi.org/10.1371/journal.pone.0014514.s006", "https://dx.doi.org/10.1371/journal.pone.0014514.s007", "https://dx.doi.org/10.1371/journal.pone.0014514.s008", "https://dx.doi.org/10.1371/journal.pone.0014514.s009", "https://dx.doi.org/10.1371/journal.pone.0014514.s010", "https://dx.doi.org/10.1371/journal.pone.0014514.s011", "https://dx.doi.org/10.1371/journal.pone.0014514.s012", "https://dx.doi.org/10.1371/journal.pone.0014514.s013", "https://dx.doi.org/10.1371/journal.pone.0014514.s014"], "stats"=>{"downloads"=>22, "page_views"=>30, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Evidence_for_OTUD_6B_Participation_in_B_Lymphocytes_Cell_Cycle_after_Cytokine_Stimulation/139515", "title"=>"Evidence for OTUD-6B Participation in B Lymphocytes Cell Cycle after Cytokine Stimulation", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-01-18 02:38:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/805714"], "description"=>"<p><b>A & B & C.</b> Real-time PCR, RT-PCR, and immunoblot of OTUD-6B, GAPDH, cyclin D1, cyclin D2, cyclin D3, p21, p27, p15, p16, cdk4, cdk6, cdc2, cyclin E, Rb, and c-Myc genes from total RNA samples and protein extracts of DOX(−) and DOX(+) Hela cells. Hela cells were provided by the Cell Bank, Shanghai Institute of Biochemistry and Cell Biology, SIBS, CAS. <b>D & E.</b> Immunoblot of OTUD-6B, cyclin D2, and p27 with gradient concentrations (0, 0.1, 0.5, 1, 2 ug/ml) of DOX stimulation. Quantitation was performed with Bandscan 5.0 from 3 independent results. <b>F & G.</b> Immunoblot (left panel) for cyclin D1, cyclin D2, and cyclin D3 on Hela and Ba/F3 cells overexpressing pcDNA3.1(+), HA-OTUD-6B WT, and HA-OTUD-6B C188S. Quantitation (right panel) of cyclin D1, cyclin D2, and cyclin D3 on Hela and Ba/F3 cells was performed with Bandscan 5.0 from three independent results.</p>", "links"=>[], "tags"=>["d2", "down-regulated", "otud-6b"], "article_id"=>476084, "categories"=>["Cell Biology"], "users"=>["Zhongping Xu", "Yufang Zheng", "Yufei Zhu", "Xiangyin Kong", "Landian Hu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014514.g004", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cyclin_D2_level_is_down_regulated_with_OTUD_6B_overexpression_/476084", "title"=>"Cyclin D2 level is down-regulated with OTUD-6B overexpression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-01-18 01:41:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/805305"], "description"=>"<p><b>A.</b> Molecular cloning of the cDNA of human OTUD-6B was amplified from total RNA of Raji cells and subjected to 1% agarose gel analysis. <b>B.</b> The expression of mouse Otud-6b mRNA in Ba/F3 cells <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0014514#pone.0014514-Zhu1\" target=\"_blank\">[5]</a>, CTLL-2 cells and various tissues (brain, heart, liver, lung, kidney, ovary, and spleen) was analyzed by RT-PCR using Otud-6b-specific primers. G3PDH was used as control. Ba/F3 cells were kindly supplied by prof. Xin yuan Liu (Shanghai Institute of Biochemistry and Cell Biology, SIBS, CAS). CTLL2 cells were provided by the Cell Bank, Shanghai Institute of Biochemistry and Cell Biology, SIBS, CAS. <b>C.</b> Ub-Met-β-gal fusion protein was prepared from MC1061 cells. The supernatant was incubated with purified GST (lane 1), OTUD-6B WT (lane 2), OTUD-6B C188S (lane 3), GST (lane 4), GST-CYLD WT (lane 5), and GST-CYLD CS (lane 6) fusion protein at 4°C with rotation for 4 hours. Both OTUD-6B WT and GST-CYLD WT could cleave ubiquitin from the Ub-Met-β-gal fusion protein.</p>", "links"=>[], "tags"=>["deubiquitinating"], "article_id"=>475673, "categories"=>["Cell Biology"], "users"=>["Zhongping Xu", "Yufang Zheng", "Yufei Zhu", "Xiangyin Kong", "Landian Hu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014514.g001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_OTUD_6B_is_a_functional_deubiquitinating_enzyme_/475673", "title"=>"OTUD-6B is a functional deubiquitinating enzyme.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-01-18 01:34:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/805396"], "description"=>"<p><b>A.</b> Otud-6b RNA level in Ba/F3 cells after starvation and two hours stimulation with different concentrations (0, 0.01, 0.1, 1, 10, 100, and 1000 pM) of mouse IL-3, IL-4, IL-13, and GM-CSF. <b>B.</b> Otud-6b RNA level in Ba/F3 cells after starvation and stimulation with 10 pM mouse IL-3, IL-4, IL-13, and GM-CSF for the indicated times (0, 0.5, 1, 2, 4, 8, and 16 hours). <b>C&D.</b> Ba/F3 cells were starved and stimulated with 10 pM mouse IL-3 or IL-4 for the indicated time. Otud-6b expression of the Ba/F3 cell lysates was immunoblotted using anti-OTUD-6B antibody. G3PDH was used as a loading control. <b>E&F.</b> Otud-6b RNA level and protein level in primary mouse B cells after starvation and stimulation with 10 pM mouse IL-3 for the indicated times (0, 0.5, 1, 2, 3, 4, 8, 16, and 24 hours). G3PDH was used as a loading control. <b>G&H.</b> Otud-6b RNA level in primary mouse B cells after starvation and two hours stimulation with different concentrations (0, 0.01, 0.1, 1, 10, 100, and 1000 pM) of mouse IL-3. G3PDH was used as a loading control.</p>", "links"=>[], "tags"=>["induced", "cytokines"], "article_id"=>475767, "categories"=>["Cell Biology"], "users"=>["Zhongping Xu", "Yufang Zheng", "Yufei Zhu", "Xiangyin Kong", "Landian Hu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014514.g002", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Otud_6b_expression_is_induced_by_cytokines_following_by_a_rapid_decline_/475767", "title"=>"Otud-6b expression is induced by cytokines following by a rapid decline.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-01-18 01:36:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/805514"], "description"=>"<p><b>A.</b> Ba/F3 cells were transfected with pcDNA3.1(+), OTUD-6B WT, and OTUD-6B C188S vectors. Thirty-two hours later, Ba/F3 cells were seeded at 2×10<sup>5</sup> cells per ml and analyzed at 16-hour intervals by trypan blue staining. <b>B.</b> Ba/F3 cells were transfected with pcDNA3.1(+), OTUD-6B WT, and OTUD-6B C188S vectors. Thirty-two hours later, cells were stained using PI and analyzed by flow cytometry. Cell cycle profile from three independent experiments was calculated statistically. <b>C.</b> PI-Annexin V analysis among pcDNA3.1(+), OTUD-6B WT, and OTUD-6B C188S-expressing Ba/F3 cells. All experiments were repeated three times independently. <b>D.</b> Ba/F3 cells were transfected with pcDNA3.1(+), OTUD-6B WT, and OTUD-6B C188S vectors. Thirty-two hours later, cell extracts were analyzed by immunoblot with anti-HA antibody. GAPDH was used as a loading control.</p>", "links"=>[], "tags"=>["overexpression", "slows", "proliferation", "increases"], "article_id"=>475877, "categories"=>["Cell Biology"], "users"=>["Zhongping Xu", "Yufang Zheng", "Yufei Zhu", "Xiangyin Kong", "Landian Hu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0014514.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_OTUD_6B_overexpression_slows_proliferation_and_increases_the_rate_of_apoptosis_/475877", "title"=>"OTUD-6B overexpression slows proliferation and increases the rate of apoptosis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-01-18 01:37:57"}

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