ZFP36L1 Negatively Regulates Plasmacytoid Differentiation of BCL1 Cells by Targeting BLIMP1 mRNA
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{"title"=>"ZFP36L1 Negatively Regulates Plasmacytoid Differentiation of BCL1 Cells by Targeting BLIMP1 mRNA", "type"=>"journal", "authors"=>[{"first_name"=>"Asghar", "last_name"=>"Nasir", "scopus_author_id"=>"55535284600"}, {"first_name"=>"John D.", "last_name"=>"Norton", "scopus_author_id"=>"7402442377"}, {"first_name"=>"Maria", "last_name"=>"Baou", "scopus_author_id"=>"19933630800"}, {"first_name"=>"Anna", "last_name"=>"Zekavati", "scopus_author_id"=>"55534294600"}, {"first_name"=>"Marie Jose", "last_name"=>"Bijlmakers", "scopus_author_id"=>"6602457170"}, {"first_name"=>"Steve", "last_name"=>"Thompson", "scopus_author_id"=>"55456071000"}, {"first_name"=>"John J.", "last_name"=>"Murphy", "scopus_author_id"=>"55678873000"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"23284928", "sgr"=>"84871416070", "doi"=>"10.1371/journal.pone.0052187", "scopus"=>"2-s2.0-84871416070", "pui"=>"366319222", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "issn"=>"19326203"}, "id"=>"3d2d4163-b05f-380e-8c70-3764ae2917a2", "abstract"=>"The ZFP36/Tis11 family of zinc-finger proteins regulate cellular processes by binding to adenine uridine rich elements in the 3' untranslated regions of various mRNAs and promoting their degradation. We show here that ZFP36L1 expression is largely extinguished during the transition from B cells to plasma cells, in a reciprocal pattern to that of ZFP36 and the plasma cell transcription factor, BLIMP1. Enforced expression of ZFP36L1 in the mouse BCL1 cell line blocked cytokine-induced differentiation while shRNA-mediated knock-down enhanced differentiation. Reconstruction of regulatory networks from microarray gene expression data using the ARACNe algorithm identified candidate mRNA targets for ZFP36L1 including BLIMP1. Genes that displayed down-regulation in plasma cells were significantly over-represented (P = <0.0001) in a set of previously validated ZFP36 targets suggesting that ZFP36L1 and ZFP36 target distinct sets of mRNAs during plasmacytoid differentiation. ShRNA-mediated knock-down of ZFP36L1 in BCL1 cells led to an increase in levels of BLIMP1 mRNA and protein, but not for mRNAs of other transcription factors that regulate plasmacytoid differentiation (xbp1, irf4, bcl6). Finally, ZFP36L1 significantly reduced the activity of a BLIMP1 3' untranslated region-driven luciferase reporter. Taken together, these findings suggest that ZFP36L1 negatively regulates plasmacytoid differentiation, at least in part, by targeting the expression of BLIMP1.", "link"=>"http://www.mendeley.com/research/zfp36l1-negatively-regulates-plasmacytoid-differentiation-bcl1-cells-targeting-blimp1-mrna", "reader_count"=>15, "reader_count_by_academic_status"=>{"Researcher"=>5, "Student > Ph. D. Student"=>7, "Student > Master"=>3}, "reader_count_by_user_role"=>{"Researcher"=>5, "Student > Ph. D. Student"=>7, "Student > Master"=>3}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>1, "Agricultural and Biological Sciences"=>9, "Medicine and Dentistry"=>2, "Immunology and Microbiology"=>3}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>9}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>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/521484"], "description"=>"<p>(A) qRT-PCR analysis of zfp36l1 mRNA levels in pSicoR.zfp36l1.RNAi1 and pSicoR.zfp36l1.RNAi2 lentivirus infected cells compared to wild-type, empty vector or pSicoR.scramble.RNAi infected cells. The results represent mean ±SD (n = 3) of zfp36l1 mRNA levels in three independent cells lines generated by three independent rounds of lentiviral infection for each cell type (apart from wild-type). * = p<0.05 as determined by t-test. (B) Western blot analysis of ZFP36L1 protein expression in wild-type, empty vector, scramble, pSicoR.scramble.RNAi1and pSicoR.scramble.RNAi2 cells. HSP90 levels are shown as a loading control.</p>", "links"=>[], "tags"=>["downregulation", "zfp36l1", "mrna", "shrna", "lentivirus", "lentiviral", "transduced", "bcl1"], "article_id"=>191968, "categories"=>["Cancer", "Molecular Biology", "Hematology", "Immunology"], "users"=>["Asghar Nasir", "John D. Norton", "Maria Baou", "Anna Zekavati", "Marie-Jose Bijlmakers", "Steve Thompson", "John J. Murphy"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052187.g004", "stats"=>{"downloads"=>2, "page_views"=>68, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effective_downregulation_of_zfp36l1_mRNA_and_protein_expression_by_zfp36l1_shRNA_lentivirus_pSicoR_zfp36l1_in_independent_lentiviral_transduced_BCL1_cell_lines_/191968", "title"=>"Effective downregulation of zfp36l1 mRNA and protein expression by zfp36l1 shRNA lentivirus (pSicoR.zfp36l1) in independent lentiviral transduced BCL1 cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-20 00:32:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/521988"], "description"=>"<p>qRT-PCR analysis of blimp1, xbp1, irf4 and bcl6 mRNA expression in pSicoR.zfp36l1.RNAi1 and pSicoR.zfp36l1.RNAi2 BCL1 cells compared to compared to wild-type, empty vector or pSicoR.scramble.RNAi infected cells. Cells were cultured in medium alone for 48h. Total RNA was extracted from 5×10<sup>6</sup> cells, 1 µg RNA was reverse transcribed and the resulting cDNA was used as template for qRT-PCR assay with mouse gene specific primers for blimp1, xbp1, irf4 and bcl6. The 2<sup>–ΔΔCT</sup> method of relative quantification was used to determine the fold change in mRNA expression compared to levels in wild-type cells. Mean ±SD (n = 3) are shown for three independent cell lines generated for each of the lentivirus infected cells. * = p<0.05 as determined by t-test.</p>", "links"=>[], "tags"=>["plasmacytoid", "differentiation", "mrnas", "bcl1"], "article_id"=>192475, "categories"=>["Cancer", "Molecular Biology", "Hematology", "Immunology"], "users"=>["Asghar Nasir", "John D. Norton", "Maria Baou", "Anna Zekavati", "Marie-Jose Bijlmakers", "Steve Thompson", "John J. Murphy"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052187.g007", "stats"=>{"downloads"=>1, "page_views"=>23, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_qRT_PCR_analysis_of_plasmacytoid_differentiation_associated_mRNAs_in_different_BCL1_cell_lines_/192475", "title"=>"qRT-PCR analysis of plasmacytoid differentiation associated mRNAs in different BCL1 cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-20 00:41:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/521419"], "description"=>"<p>IgM production. Levels of IgM production in ZFP36L1 transfected BCL1 stimulated with IL-2/5 (20 ng/ml IL-2 and 5 ng/ml IL-5) over 4 days compared to levels produced by empty vector BCL1 cells cultured under the same conditions. Cells were co-transfected with either pcDNA3.ZFP36L1 or empty pcDNA3 vector and pcDNA3.EGFP and then sorted on the basis of EGFP expression before been set up in culture in the absence or presence of cytokines. Cells were cultured at 4.0×10<sup>4</sup>/ml and ELISA measurements were made in duplicate. The results shown are representative of 3 similar experiments.</p>", "links"=>[], "tags"=>["zfp36l1", "bcl1", "cells", "inhibits"], "article_id"=>191908, "categories"=>["Cancer", "Molecular Biology", "Hematology", "Immunology"], "users"=>["Asghar Nasir", "John D. Norton", "Maria Baou", "Anna Zekavati", "Marie-Jose Bijlmakers", "Steve Thompson", "John J. Murphy"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052187.g003", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ectopic_expression_of_ZFP36L1_in_BCL1_cells_inhibits_cytokine_induced_/191908", "title"=>"Ectopic expression of ZFP36L1 in BCL1 cells inhibits cytokine-induced.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-20 00:31:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/521801"], "description"=>"<p>(A) Heat map expression profile of known ZFP36L1 target genes in terminal B cell differentiation. The profiles are compared with the reference genes, ZFP36L1, BLIMP1 and XBP1 (first three rows). GMCSF, VEGFA and IL-3 are validated ZFP36L1 targets; the remaining genes have been validated for ZFP36 <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0052187#pone.0052187-Baou1\" target=\"_blank\">[4]</a>. Data was taken from GEO Accession number GSE 6691 <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0052187#pone.0052187-Gutierrez1\" target=\"_blank\">[29]</a>. Red indicates high and blue, low expression. Key: CLL: B chronic lymphocytic leukemia, MM: multiple myeloma, WM-BL: Waldenstrom’s macroglobulinemia B cells, WM-PB: Waldenstrom’s macroglobulinemia plasma cells, NBC: normal B cells, PC: normal plasma cells. (B) Graphical representation of the ARACNe network for BLIMP1 and ZFP36L1. Nodes representing the BLIMP1 and ZFP36L1 hubs are shown enlarged. Only first neighbours of these hubs are shown in the module. Nodes representing inferred BLIMP1 targets (significantly up-regulated in normal B cells) are blue; nodes representing inferred ZFP36L1 targets (significantly up-regulated in normal plasma cells) are red. Network graphics were generated as group attribute layout using Cytoscape version 2.6.0. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0052187#pone.0052187-Cline1\" target=\"_blank\">[35]</a>.</p>", "links"=>[], "tags"=>["aracne"], "article_id"=>192295, "categories"=>["Cancer", "Molecular Biology", "Hematology", "Immunology"], "users"=>["Asghar Nasir", "John D. Norton", "Maria Baou", "Anna Zekavati", "Marie-Jose Bijlmakers", "Steve Thompson", "John J. Murphy"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052187.g006", "stats"=>{"downloads"=>1, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Microarray_expression_profile_and_ARACNe_network_of_ZFP36L1_/192295", "title"=>"Microarray expression profile and ARACNe network of ZFP36L1.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-20 00:38:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/281282", "https://ndownloader.figshare.com/files/281332", "https://ndownloader.figshare.com/files/281383", "https://ndownloader.figshare.com/files/281424", "https://ndownloader.figshare.com/files/281515", "https://ndownloader.figshare.com/files/281570", "https://ndownloader.figshare.com/files/281656", "https://ndownloader.figshare.com/files/281728"], "description"=>"<div><p>The ZFP36/Tis11 family of zinc-finger proteins regulate cellular processes by binding to adenine uridine rich elements in the 3′ untranslated regions of various mRNAs and promoting their degradation. We show here that ZFP36L1 expression is largely extinguished during the transition from B cells to plasma cells, in a reciprocal pattern to that of ZFP36 and the plasma cell transcription factor, BLIMP1. Enforced expression of ZFP36L1 in the mouse BCL1 cell line blocked cytokine-induced differentiation while shRNA-mediated knock-down enhanced differentiation. Reconstruction of regulatory networks from microarray gene expression data using the ARACNe algorithm identified candidate mRNA targets for ZFP36L1 including BLIMP1. Genes that displayed down-regulation in plasma cells were significantly over-represented (P = <0.0001) in a set of previously validated ZFP36 targets suggesting that ZFP36L1 and ZFP36 target distinct sets of mRNAs during plasmacytoid differentiation. ShRNA-mediated knock-down of ZFP36L1 in BCL1 cells led to an increase in levels of BLIMP1 mRNA and protein, but not for mRNAs of other transcription factors that regulate plasmacytoid differentiation (xbp1, irf4, bcl6). Finally, ZFP36L1 significantly reduced the activity of a BLIMP1 3′ untranslated region-driven luciferase reporter. Taken together, these findings suggest that ZFP36L1 negatively regulates plasmacytoid differentiation, at least in part, by targeting the expression of BLIMP1.</p> </div>", "links"=>[], "tags"=>["zfp36l1", "negatively", "regulates", "plasmacytoid", "differentiation", "bcl1", "cells", "targeting", "blimp1", "mrna"], "article_id"=>115465, "categories"=>["Cancer", "Molecular Biology", "Hematology", "Immunology"], "users"=>["Asghar Nasir", "John D. Norton", "Maria Baou", "Anna Zekavati", "Marie-Jose Bijlmakers", "Steve Thompson", "John J. Murphy"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0052187.s001", "https://dx.doi.org/10.1371/journal.pone.0052187.s002", "https://dx.doi.org/10.1371/journal.pone.0052187.s003", "https://dx.doi.org/10.1371/journal.pone.0052187.s004", "https://dx.doi.org/10.1371/journal.pone.0052187.s005", "https://dx.doi.org/10.1371/journal.pone.0052187.s006", "https://dx.doi.org/10.1371/journal.pone.0052187.s007", "https://dx.doi.org/10.1371/journal.pone.0052187.s008"], "stats"=>{"downloads"=>14, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/ZFP36L1_Negatively_Regulates_Plasmacytoid_Differentiation_of_BCL1_Cells_by_Targeting_BLIMP1_mRNA__/115465", "title"=>"ZFP36L1 Negatively Regulates Plasmacytoid Differentiation of BCL1 Cells by Targeting BLIMP1 mRNA", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-12-20 01:31:05"}
  • {"files"=>["https://ndownloader.figshare.com/files/521300"], "description"=>"<p><b>downregulation of ZFP36L1 is associated with plasmacytoid differentiation of B cells.</b> (A) Western Blot analysis of ZFP36L1 expression in IL-2/5 treated murine leukemic BCL1 cells. Protein lysates were made from unstimulated cells (lane 1) or from cells 96 h after stimulation with cytokines (20 ng/ml IL-2 and 5 ng/ml IL-5) (lane 2). ZFP36L1 and HSP90 proteins were detected by anti-BRF1/2 and anti-HSP 90 antibodies respectively. (B) qRT-PCR analysis of zfp36l1 and blimp1 mRNA expression in day 0 versus 48 h IL-2/5 stimulated BCL1 cells. (C) qRT-PCR analysis of time course of zfp36l1 and blimp1 mRNA expression over 3 days in LPS (10 µg/ml) stimulated murine splenic B cells. The 2<sup>–ΔΔCT</sup> method of relative quantification was used to determine the fold change in mRNA expression. The results shown were normalized to β-actin mRNA expression. The results show mean ±SD from one representative experiment.</p>", "links"=>[], "tags"=>["immunology", "molecular biology", "hematology", "oncology"], "article_id"=>191790, "categories"=>["Cancer", "Molecular Biology", "Hematology", "Immunology"], "users"=>["Asghar Nasir", "John D. Norton", "Maria Baou", "Anna Zekavati", "Marie-Jose Bijlmakers", "Steve Thompson", "John J. Murphy"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052187.g002", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Significant_/191790", "title"=>"Significant", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-20 00:29:50"}
  • {"files"=>["https://ndownloader.figshare.com/files/522117"], "description"=>"<p>(A) Western blot analysis of BLIMP1 levels in control and ZFP36L1 knockdown cells. BLIMP1 expression levels are upregulated in ZFP36L1 knockdown cells compared to controls. βACTIN levels are shown as a loading control. (B) ZFP36L1 mediates degradation of the BLIMP1 3′UTR. HEK 293 T cells were transfected with pMIRBLIMP1 3′UTR construct alone (control) or with either ZFP36L1 or a zinc finger domain mutant, ZFP36L1 Mut. Renilla luciferase was also included in all transfections as a normalization control. 24 hours later cell lysates were harvested and firefly and renilla luciferase levels measured using a Fluorstar Optima plate reader. Relative levels of luciferase activity were measured. Mean ±SD, are shown for four independent experiments (n = 4), * = p<0.05 as determined by t-test.</p>", "links"=>[], "tags"=>["knockdown", "bcl1", "cells", "blimp1", "upregulation", "zfp36l1", "interacts", "blimp"], "article_id"=>192593, "categories"=>["Cancer", "Molecular Biology", "Hematology", "Immunology"], "users"=>["Asghar Nasir", "John D. Norton", "Maria Baou", "Anna Zekavati", "Marie-Jose Bijlmakers", "Steve Thompson", "John J. Murphy"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052187.g008", "stats"=>{"downloads"=>2, "page_views"=>29, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_ZFP36L1_knockdown_in_BCL1_cells_is_associated_with_BLIMP1_upregulation_and_ZFP36L1_interacts_with_the_BLIMP_3_8242_UTR_/192593", "title"=>"ZFP36L1 knockdown in BCL1 cells is associated with BLIMP1 upregulation and ZFP36L1 interacts with the BLIMP 3′UTR.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-20 00:43:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/521608"], "description"=>"<p>Cell numbers in the absence (A) and presence (B) of cytokines (IL-2 and IL-5) in pSicoR.zfp36l1.RNAi1 and pSicoR.zfp36l1.RNAi2 BCL1 cells compared to compared to wild-type, empty vector or pSicoR.scramble.RNAi infected cells. Cells numbers were counted in quadruplicate 4 days after seeding 2×10<sup>5</sup>/ml cells on day 0 in flasks in the absence or presence of cytokines (IL-2 20 ng/ml and IL-5 5 ng/ml). Mean ±SD (n = 3) are shown for three independent cell lines generated for each of the lentivirus infected cells. IgM secretion (ng/ml) measured by ELISA, 4 days after the start of the culture, in the absence (C) and presence (D) of cytokines (IL-2 20 ng/ml and IL-5 5 ng/ml) in pSicoR.zfp36l1.RNAi1 and pSicoR.zfp36l1.RNAi2 BCL1 cells compared to compared to wild-type, empty vector or pSicoR.scramble.RNAi infected cells. Mean ±SD (n = 3) are shown for three independent cell lines generated for each of the lentivirus infected cells. ** = p<0.01, * = p<0.05 as determined by t-test.</p>", "links"=>[], "tags"=>["knockdown", "zfp36l1", "bcl1", "cells", "plasmacytoid", "differentiation", "inducing", "igm"], "article_id"=>192091, "categories"=>["Cancer", "Molecular Biology", "Hematology", "Immunology"], "users"=>["Asghar Nasir", "John D. Norton", "Maria Baou", "Anna Zekavati", "Marie-Jose Bijlmakers", "Steve Thompson", "John J. Murphy"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052187.g005", "stats"=>{"downloads"=>3, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Lentiviral_mediated_knockdown_of_ZFP36L1_expression_in_BCL1_cells_promotes_plasmacytoid_differentiation_by_inducing_IgM_production_/192091", "title"=>"Lentiviral-mediated knockdown of ZFP36L1 expression in BCL1 cells promotes plasmacytoid differentiation by inducing IgM production.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-20 00:34:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/521217"], "description"=>"<p>(A) RT-PCR analysis of ZFP36L1 expression in human malignant B cells representing different stages of B cell differentiation. For comparison BLIMP1 expression was also measured. β ACTIN expression levels are shown as a loading control. (B) Comparison of ZFP36L1 protein levels in human Ramos B cells before and after 3 h PMA stimulation and in two myeloma cell lines RPMI-8226 cells and KMS-11. BLIMP1 expression is also shown in RPMI-8226 cells and KMS-11 cells. HSP90 levels are shown as a loading control. ZFP36L1/L2, HSP90 and BLIMP1 proteins were detected by anti-BRF1/2, anti-HSP90 and anti-BLIMP1 antibodies respectively. Anti-BRF1/2 antibody cross-reacts with ZFP36L2 (approx. 60 kDa) and ZFP36L1 appears typically as a constellation of induced bands (approx. 40 kDa) in human cells.</p>", "links"=>[], "tags"=>["cells", "stages"], "article_id"=>191708, "categories"=>["Cancer", "Molecular Biology", "Hematology", "Immunology"], "users"=>["Asghar Nasir", "John D. Norton", "Maria Baou", "Anna Zekavati", "Marie-Jose Bijlmakers", "Steve Thompson", "John J. Murphy"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052187.g001", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_ZFP36L1_expression_in_human_B_cells_at_various_stages_of_differentiation_/191708", "title"=>"ZFP36L1 expression in human B cells at various stages of differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-20 00:28:28"}

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Relative Metric

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