Characterisation of Translation Elongation Factor eEF1B Subunit Expression in Mammalian Cells and Tissues and Co-Localisation with eEF1A2
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{"title"=>"Characterisation of translation elongation factor eEF1B subunit expression in mammalian cells and tissues and co-localisation with eEF1A2", "type"=>"journal", "authors"=>[{"first_name"=>"Yuan", "last_name"=>"Cao", "scopus_author_id"=>"56437039900"}, {"first_name"=>"Miriam", "last_name"=>"Portela", "scopus_author_id"=>"7006833995"}, {"first_name"=>"Justyna", "last_name"=>"Janikiewicz", "scopus_author_id"=>"57195150027"}, {"first_name"=>"Jennifer", "last_name"=>"Doig", "scopus_author_id"=>"7003658234"}, {"first_name"=>"Catherine M.", "last_name"=>"Abbott", "scopus_author_id"=>"7102454330"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84914689162", "scopus"=>"2-s2.0-84914689162", "doi"=>"10.1371/journal.pone.0114117", "pui"=>"600627909", "issn"=>"19326203", "pmid"=>"25436608"}, "id"=>"2405b6b2-3c15-300b-9dbf-141421202faf", "abstract"=>"Translation elongation is the stage of protein synthesis in which the translation factor eEF1A plays a pivotal role that is dependent on GTP exchange. In vertebrates, eEF1A can exist as two separately encoded tissue-specific isoforms, eEF1A1, which is almost ubiquitously expressed, and eEF1A2, which is confined to neurons and muscle. The GTP exchange factor for eEF1A1 is a complex called eEF1B made up of subunits eEF1Bα, eEF1Bδ and eEF1Bγ. Previous studies have cast doubt on the ability of eEF1B to interact with eEF1A2, suggesting that this isoform might use a different GTP exchange factor. We show that eEF1B subunits are all widely expressed to varying degrees in different cell lines and tissues, and at different stages of development. We show that ablation of any of the subunits in human cell lines has a small but significant impact on cell viability and cycling. Finally, we show that both eEF1A1 and eEF1A2 colocalise with all eEF1B subunits, in such close proximity that they are highly likely to be in a complex.", "link"=>"http://www.mendeley.com/research/characterisation-translation-elongation-factor-eef1b-subunit-expression-mammalian-cells-tissues-colo", "reader_count"=>15, "reader_count_by_academic_status"=>{"Student > Doctoral Student"=>3, "Researcher"=>3, "Student > Ph. D. Student"=>3, "Student > Master"=>1, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_user_role"=>{"Student > Doctoral Student"=>3, "Researcher"=>3, "Student > Ph. D. Student"=>3, "Student > Master"=>1, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>3, "Medicine and Dentistry"=>3, "Agricultural and Biological Sciences"=>5, "Neuroscience"=>1, "Chemistry"=>3}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>5}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1811739"], "description"=>"<p>Panel A: Immunoblot of eEF1Bα, eEF1Bδ and eEF1Bγ protein expression in cell lines. GAPDH was used as a loading control. Panel B: Immunoblot of eEF1B subunits in 24-day old mice. Each tissue is shown twice, the left hand sample is from a wild type mouse and the right hand sample from a wasted homozygous mouse. SC =  spinal cord. The second row shows a longer exposure of the eEF1Bδ blot, revealing the expression of the longer isoform. GAPDH was used as a loading control. Panel C: Immunoblot of eEF1Bδ in an extended panel of mouse tissues showing brain and testis-expression of the longer isoform (top panel shows a longer exposure). GAPDH was used as a loading control. Panel D: Immunoblot of eEF1B subunits expression in brain (Br) and liver (Li) throughout late embryonic and postnatal mouse development. GAPDH was used as a loading control.</p>", "links"=>[], "tags"=>["translation factor eEF 1A", "eEF 1B subunits", "subunits eEF 1B eEF 1B", "GTP exchange factor", "eEF 1A Translation elongation", "eEF 1A colocalise", "Cell lines", "eEF 1A", "Translation Elongation Factor eEF 1B Subunit Expression", "eEF 1B"], "article_id"=>1255982, "categories"=>["Biological Sciences"], "users"=>["Yuan Cao", "Miriam Portela", "Justyna Janikiewicz", "Jennifer Doig", "Catherine M. Abbott"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114117.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_analysis_of_eEF1B_subunits_in_cell_lines_and_tissues_/1255982", "title"=>"Expression analysis of eEF1B subunits in cell lines and tissues.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-01 03:34:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/1811752"], "description"=>"<p>Immunohistochemistry of eEF1B subunits in human and mouse brain and pancreas. Proteins detected through primary antibody incubation, HRP mouse + rabbit secondary antibody and subsequent incubation with DAB. Positive signal is indicated by the presence of brown DAB reaction product. Panels show eEF1Bα (<b>b</b> and <b>f</b>), eEF1Bδ(<b>c</b> and <b>g</b>) and eEF1Bγ (<b>d</b> and <b>h</b>) Incubation with secondary antibody only was used as negative control (<b>a</b> and <b>e</b>). Bar (top left micrograph) represents 100 and 50 µm respectively.</p>", "links"=>[], "tags"=>["translation factor eEF 1A", "eEF 1B subunits", "subunits eEF 1B eEF 1B", "GTP exchange factor", "eEF 1A Translation elongation", "eEF 1A colocalise", "Cell lines", "eEF 1A", "Translation Elongation Factor eEF 1B Subunit Expression", "eEF 1B"], "article_id"=>1255995, "categories"=>["Biological Sciences"], "users"=>["Yuan Cao", "Miriam Portela", "Justyna Janikiewicz", "Jennifer Doig", "Catherine M. Abbott"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114117.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemistry_of_eEF1B_subunits_in_human_and_mouse_tissues_/1255995", "title"=>"Immunohistochemistry of eEF1B subunits in human and mouse tissues.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-01 03:34:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/1811753"], "description"=>"<p>Immunoblots of protein extracts from cell lines after RNA interference. Panel A: eEF1Bα, eEF1Bδ and eEF1Bγ protein level efficiently knocked down by three different siRNAs in HeLa cells 72 h after transfection. GAPDH was used as a loading control. Panel B: eEF1Bα, eEF1Bδ and eEF1Bγ protein level efficiently knocked down by three different siRNAs in HCT116 and DLD1 cells 72 h after transfection. GAPDH was used as a loading control.</p>", "links"=>[], "tags"=>["translation factor eEF 1A", "eEF 1B subunits", "subunits eEF 1B eEF 1B", "GTP exchange factor", "eEF 1A Translation elongation", "eEF 1A colocalise", "Cell lines", "eEF 1A", "Translation Elongation Factor eEF 1B Subunit Expression", "eEF 1B"], "article_id"=>1255996, "categories"=>["Biological Sciences"], "users"=>["Yuan Cao", "Miriam Portela", "Justyna Janikiewicz", "Jennifer Doig", "Catherine M. Abbott"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114117.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Using_RNA_interference_to_knock_down_expression_of_eEF1B_subunits_/1255996", "title"=>"Using RNA interference to knock down expression of eEF1B subunits.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-01 03:34:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/1811759"], "description"=>"<p>Panel A: A decrease in cellular metabolism is observed when any of the eEF1B subunit protein level is decreased by siRNAs in HeLa, HTC116 and DLD1 cells. Cell metabolism was assessed by the Alamar blue assay. Data were obtained from the mean of three or more independent experiments in HeLa, HCT116 and DLD1 cells, with more than 10 wells each. Error bars indicate +- SEM; n of wells>10; n = 3−4; *, P<0.05; ***, P<0.001 from non-targeting siRNA. Panel B: Knockdown of eEF1B subunits leads to altered cell cycle profile in three cell lines: representative images of the flow cytometry analysis. Error bars indicate +- SEM; n = 3; *, P<0.05; **, P<0.01 of non-targeting siRNA.</p>", "links"=>[], "tags"=>["translation factor eEF 1A", "eEF 1B subunits", "subunits eEF 1B eEF 1B", "GTP exchange factor", "eEF 1A Translation elongation", "eEF 1A colocalise", "Cell lines", "eEF 1A", "Translation Elongation Factor eEF 1B Subunit Expression", "eEF 1B"], "article_id"=>1256002, "categories"=>["Biological Sciences"], "users"=>["Yuan Cao", "Miriam Portela", "Justyna Janikiewicz", "Jennifer Doig", "Catherine M. Abbott"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114117.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Viability_and_cell_cycle_distribution_of_cells_after_ablation_of_eEF1B_subunits_/1256002", "title"=>"Viability and cell cycle distribution of cells after ablation of eEF1B subunits.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-01 03:34:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/1811767"], "description"=>"<p>IF images of the expression of eEF1A2 and eEF1Bα (top panel) or eEF1Bδ (bottom panel) on mouse spinal cord.</p>", "links"=>[], "tags"=>["translation factor eEF 1A", "eEF 1B subunits", "subunits eEF 1B eEF 1B", "GTP exchange factor", "eEF 1A Translation elongation", "eEF 1A colocalise", "Cell lines", "eEF 1A", "Translation Elongation Factor eEF 1B Subunit Expression", "eEF 1B"], "article_id"=>1256010, "categories"=>["Biological Sciences"], "users"=>["Yuan Cao", "Miriam Portela", "Justyna Janikiewicz", "Jennifer Doig", "Catherine M. Abbott"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114117.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_in_mouse_spinal_cord_/1256010", "title"=>"Expression in mouse spinal cord.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-01 03:34:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/1811775"], "description"=>"<p>Panel I: PLA on HeLa cells. A. Negative control with both primary antibodies omitted. B. eEF1A2 antibody only. C. PLA of eEF1A2 and eEF1Bα. D. PLA of eEF1A2 and eEF1Bδ. E. PLA of eEF1A2 and eEF1Bγ. F. PLA of eEF1A2 and TK1 as negative control. Images in the squares are higher magnification of selected areas. Panel II: PLA on NIH-3T3 cells stably expressing V5-tagged eEF1A1. A. Negative control with both primary antibodies omitted. B. V5 antibody only. C. PLA of eEF1Bα and V5. D. PLA of eEF1Bα and TK1. E. PLA of eEF1Bδ and V5. F. PLA of eEF1Bδ and TK1. G. PLA of eEF1Bγ and V5. H. PLA of eEF1Bγ and TK1. Images in the squares are higher magnification of selected areas. Panel III: PLA on NIH-3T3 cells stably expressing V5-tagged eEF1A2. A. Negative control with both primary antibodies omitted. B. V5 antibody only. C. PLA of eEF1Bα and V5. D. PLA of eEF1Bα and TK1. E. PLA of eEF1Bδ and V5. F. PLA of eEF1Bδ and TK1. G. PLA of eEF1Bγ and V5. H. PLA of eEF1Bγ and TK1. Images in the squares are higher magnification of selected areas.</p>", "links"=>[], "tags"=>["translation factor eEF 1A", "eEF 1B subunits", "subunits eEF 1B eEF 1B", "GTP exchange factor", "eEF 1A Translation elongation", "eEF 1A colocalise", "Cell lines", "eEF 1A", "Translation Elongation Factor eEF 1B Subunit Expression", "eEF 1B"], "article_id"=>1256018, "categories"=>["Biological Sciences"], "users"=>["Yuan Cao", "Miriam Portela", "Justyna Janikiewicz", "Jennifer Doig", "Catherine M. Abbott"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114117.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Proximity_ligation_assays_for_eEF1A_and_eEF1B_/1256018", "title"=>"Proximity ligation assays for eEF1A and eEF1B.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-01 03:34:17"}

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