Interference in DNA Replication Can Cause Mitotic Chromosomal Breakage Unassociated with Double-Strand Breaks
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{"title"=>"Interference in DNA Replication Can Cause Mitotic Chromosomal Breakage Unassociated with Double-Strand Breaks", "type"=>"journal", "authors"=>[{"first_name"=>"Mari", "last_name"=>"Fujita", "scopus_author_id"=>"55638687900"}, {"first_name"=>"Hiroyuki", "last_name"=>"Sasanuma", "scopus_author_id"=>"16176402500"}, {"first_name"=>"Kimiyo N.", "last_name"=>"Yamamoto", "scopus_author_id"=>"35072125300"}, {"first_name"=>"Hiroshi", "last_name"=>"Harada", "scopus_author_id"=>"55127694300"}, {"first_name"=>"Aya", "last_name"=>"Kurosawa", "scopus_author_id"=>"36801376300"}, {"first_name"=>"Noritaka", "last_name"=>"Adachi", "scopus_author_id"=>"7201601863"}, {"first_name"=>"Masato", "last_name"=>"Omura", "scopus_author_id"=>"55638511400"}, {"first_name"=>"Masahiro", "last_name"=>"Hiraoka", "scopus_author_id"=>"55160393000"}, {"first_name"=>"Shunichi", "last_name"=>"Takeda", "scopus_author_id"=>"35428932700"}, {"first_name"=>"Kouji", "last_name"=>"Hirota", "scopus_author_id"=>"7402829776"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84875702315", "pui"=>"368649202", "doi"=>"10.1371/journal.pone.0060043", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"84875702315", "pmid"=>"23573231"}, "id"=>"96adb8ba-c6e0-3d22-8bee-7e8c80a00cb1", "abstract"=>"Morphological analysis of mitotic chromosomes is used to detect mutagenic chemical compounds and to estimate the dose of ionizing radiation to be administered. It has long been believed that chromosomal breaks are always associated with double-strand breaks (DSBs). We here provide compelling evidence against this canonical theory. We employed a genetic approach using two cell lines, chicken DT40 and human Nalm-6. We measured the number of chromosomal breaks induced by three replication-blocking agents (aphidicolin, 5-fluorouracil, and hydroxyurea) in DSB-repair-proficient wild-type cells and cells deficient in both homologous recombination and nonhomologous end-joining (the two major DSB-repair pathways). Exposure of cells to the three replication-blocking agents for at least two cell cycles resulted in comparable numbers of chromosomal breaks for RAD54(-/-/)KU70(-/-) DT40 clones and wild-type cells. Likewise, the numbers of chromosomal breaks induced in RAD54(-/-/)LIG4(-/-) Nalm-6 clones and wild-type cells were also comparable. These data indicate that the replication-blocking agents can cause chromosomal breaks unassociated with DSBs. In contrast with DSB-repair-deficient cells, chicken DT40 cells deficient in PIF1 or ATRIP, which molecules contribute to the completion of DNA replication, displayed higher numbers of mitotic chromosomal breaks induced by aphidicolin than did wild-type cells, suggesting that single-strand gaps left unreplicated may result in mitotic chromosomal breaks.", "link"=>"http://www.mendeley.com/research/interference-dna-replication-cause-mitotic-chromosomal-breakage-unassociated-doublestrand-breaks", "reader_count"=>24, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>4, "Researcher"=>6, "Student > Ph. D. Student"=>10, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>4, "Researcher"=>6, "Student > Ph. D. Student"=>10, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>4, "Medicine and Dentistry"=>3, "Agricultural and Biological Sciences"=>14}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>14}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>3}}, "reader_count_by_country"=>{"Canada"=>1, "Japan"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1009269"], "description"=>"<p>(A–D) Frequency of chromosomal aberrations (CAs) in <i>wild-type</i> and <i>RAD54<sup>−/−/</sup>KU70<sup>−/−</sup></i> DT40 cells before (0) and after treatment with (A) γ-irradiation, (B) 5-FU, (C) HU, and (D) aphidicolin (APH). Cells were analyzed at 3 h after irradiation (A). Cells were incubated with 5-FU or HU for 24 h, or with aphidicolin for 48 h at the indicated concentrations (B–D). In each case, cells were treated with colcemid for the last 3 h. More than 100 cells were analyzed in each case. (E–H) Frequency of chromosomal aberrations (CAs) in <i>wild-type</i> and <i>RAD54<sup>−/−/</sup>LIG4<sup>−/−</sup></i> Nalm-6 cells before (0) and after treatment with (E) γ-irradiation, (F) 5-FU, and (G) HU. Cells were incubated with 5-FU or HU for 48 h at the indicated concentrations (F, G). (H) The number of induced CAs was calculated by subtracting the number of non-treated cells from the number of cells treated with γ-rays or chemicals. More than 50 cells were analyzed at 8 h after irradiation at 0.1 Gy. More than 100 cells were analyzed for 5-FU and HU. Error bars show standard error, based on the Poisson distribution of spontaneous chromosomal aberrations observed previously <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060043#pone.0060043-Sonoda2\" target=\"_blank\">[37]</a>, <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060043#pone.0060043-Kikuchi1\" target=\"_blank\">[52]</a>.</p>", "links"=>[], "tags"=>["agents", "induce", "numbers", "chromosome", "breaks", "dsb-repair-proficient", "-deficient", "dt40", "nalm-6"], "article_id"=>669869, "categories"=>["Molecular Biology", "Biochemistry", "Cancer", "Cell Biology", "Genetics"], "users"=>["Mari Fujita", "Hiroyuki Sasanuma", "Kimiyo N. Yamamoto", "Hiroshi Harada", "Aya Kurosawa", "Noritaka Adachi", "Masato Omura", "Masahiro Hiraoka", "Shunichi Takeda", "Kouji Hirota"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060043.g002", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Replication_blocking_agents_induce_comparable_numbers_of_chromosome_breaks_in_both_DSB_repair_proficient_and_deficient_chicken_DT40_and_human_Nalm_6_cell_lines_/669869", "title"=>"Replication-blocking agents induce comparable numbers of chromosome breaks in both DSB-repair-proficient and -deficient chicken DT40 and human Nalm-6 cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-03 02:44:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/1009268"], "description"=>"<p>(A) Indicated cells were either irradiated with γ-rays and cultured for 48 h or continuously incubated with aphidicolin for 72 h or, with 5-FU or HU for 48 h. Living cells were measured in terms of level of cellular ATP. The average for three independent experiments is shown. Error bars show the standard deviation for three independent experiments. (B) Cell-cycle analysis of <i>wild-type</i> DT40 cells after treatment for 4 h and 24 h with 45 µM 5-FU, 25 µM HU, and 0.25 µM aphidicolin (APH). The x-axis represents the intensity of PI staining (linear scale) and the y-axis represents bromodeoxyuridine (BrdU) uptake during 10 min pulse-labeling (logarithmic scale). The BrdU-positive fraction defined by the square was quantified. (C) Growth curves for <i>wild-type</i> DT40 and <i>RAD54<sup>−/−/</sup>KU70<sup>−/−</sup></i> cells over 0 to 30 h exposure to 0.25 µM aphidicolin, 45 µM 5-FU, and 25 µM HU. The number of live cells was counted every 6 h. The average of three independent experiments is shown. Error bars show the standard deviation for three independent experiments.</p>", "links"=>[], "tags"=>["aphidicolin", "dt40"], "article_id"=>669868, "categories"=>["Molecular Biology", "Biochemistry", "Cancer", "Cell Biology", "Genetics"], "users"=>["Mari Fujita", "Hiroyuki Sasanuma", "Kimiyo N. Yamamoto", "Hiroshi Harada", "Aya Kurosawa", "Noritaka Adachi", "Masato Omura", "Masahiro Hiraoka", "Shunichi Takeda", "Kouji Hirota"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060043.g001", "stats"=>{"downloads"=>1, "page_views"=>40, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparable_sensitivity_to_5_FU_HU_and_aphidicolin_for_wild_type_and_RAD54_8722_8722_KU70_8722_8722_DT40_cells_/669868", "title"=>"Comparable sensitivity to 5-FU, HU, and aphidicolin for <i>wild-type</i> and <i>RAD54<sup>−/−/</sup>KU70<sup>−/−</sup></i> DT40 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-03 02:44:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1009276"], "description"=>"<p>(A) Cells with the indicated genotype were exposed to the indicated replication-blocking agents and DNA damage agents. The dose of the agents is displayed on the x-axis on a linear scale, while the percent fraction of surviving cells is displayed on the y-axis on a logarithmic scale. Error bars show standard deviation of mean for three independent assays. (B) Frequency of chromosomal aberrations (CAs) in <i>wild-type</i>, <i>PIF1<sup>−/−</sup></i>, and <i>RAD54<sup>−/−/</sup>KU70<sup>−/−</sup></i> DT40 cells before (0) and after treatment with aphidicolin at indicated concentration for 48 h. (C) Percentage of the cells carrying the indicated number of chromosomal breaks is indicated as a histogram. Indicated cells were treated with 0.1 µM aphidicolin (APH) for 24 h. More than 50 cells were analyzed in each case. Error bars show standard error for the number of CAs in 50 mitotic cells, calculated as in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060043#pone-0060043-g002\" target=\"_blank\">Figure 2</a>.</p>", "links"=>[], "tags"=>["pif1", "atrip", "chromosomal"], "article_id"=>669876, "categories"=>["Molecular Biology", "Biochemistry", "Cancer", "Cell Biology", "Genetics"], "users"=>["Mari Fujita", "Hiroyuki Sasanuma", "Kimiyo N. Yamamoto", "Hiroshi Harada", "Aya Kurosawa", "Noritaka Adachi", "Masato Omura", "Masahiro Hiraoka", "Shunichi Takeda", "Kouji Hirota"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060043.g005", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Contribution_of_PIF1_and_ATRIP_to_the_prevention_of_chromosomal_breakage_/669876", "title"=>"Contribution of PIF1 and ATRIP to the prevention of chromosomal breakage.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-03 02:44:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1009273"], "description"=>"<p>(A) Cell-cycle analysis after treatment with 2 mM HU for 2 h. The BrdU-positive fraction was quantified as in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060043#pone-0060043-g001\" target=\"_blank\">Figure 1</a>. (B) Frequency of chromosomal aberrations (CAs) for <i>wild-type</i> DT40 cells and <i>RAD54<sup>−/−/</sup>KU70<sup>−/−</sup></i> cells. Cells were exposed to 2 mM HU for 2 h and then released in a drug-free medium for 3 or 6 h. (C) Cell-cycle analysis after treatment for 2 h with 0.5 µM aphidicolin (APH). The BrdU-positive fraction was quantified as in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060043#pone-0060043-g001\" target=\"_blank\">Figure 1</a>. (D) Frequency of chromosomal aberrations (CAs) for <i>wild-type</i> DT40 cells and <i>RAD54<sup>−/−/</sup>KU70<sup>−/−</sup></i> cells. Cells were exposed with 0.5 µM aphidicolin (APH) for 2 h and released in a drug-free medium for 3 or 6 h. More than 50 cells were analyzed in each case. Error bars show standard error for the number of CAs in 50 mitotic cells, calculated as in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060043#pone-0060043-g002\" target=\"_blank\">Figure 2</a>. Asterisk and double asterisk: significant difference compared with <i>wild-type cells</i> (P<0.05).</p>", "links"=>[], "tags"=>["replication", "blockage", "aphidicolin", "hu"], "article_id"=>669873, "categories"=>["Molecular Biology", "Biochemistry", "Cancer", "Cell Biology", "Genetics"], "users"=>["Mari Fujita", "Hiroyuki Sasanuma", "Kimiyo N. Yamamoto", "Hiroshi Harada", "Aya Kurosawa", "Noritaka Adachi", "Masato Omura", "Masahiro Hiraoka", "Shunichi Takeda", "Kouji Hirota"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060043.g004", "stats"=>{"downloads"=>1, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Release_from_complete_replication_blockage_by_a_high_concentration_of_aphidicolin_or_HU_induces_DSBs_/669873", "title"=>"Release from complete replication blockage by a high concentration of aphidicolin or HU induces DSBs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-03 02:44:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/1009271"], "description"=>"<p>Cells were exposed to 0.25 µM aphidicolin (APH) for 48 h. Frequency of chromosomal aberrations (CAs) for <i>wild-type</i> DT40 cells and <i>RAD54<sup>−/−/</sup>KU70<sup>−/−</sup></i> cells is shown. More than 50 cells were analyzed in each case. Error bars show the standard error for the number of CAs in 50 mitotic cells, calculated as in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060043#pone-0060043-g002\" target=\"_blank\">Figure 2</a>. In each case, cells were incubated with colcemid for the last 3 h.</p>", "links"=>[], "tags"=>["chromosomal", "breakage", "replication-blocking"], "article_id"=>669871, "categories"=>["Molecular Biology", "Biochemistry", "Cancer", "Cell Biology", "Genetics"], "users"=>["Mari Fujita", "Hiroyuki Sasanuma", "Kimiyo N. Yamamoto", "Hiroshi Harada", "Aya Kurosawa", "Noritaka Adachi", "Masato Omura", "Masahiro Hiraoka", "Shunichi Takeda", "Kouji Hirota"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060043.g003", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparable_chromosomal_breakage_after_removal_of_replication_blocking_agents_/669871", "title"=>"Comparable chromosomal breakage after removal of replication-blocking agents.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-03 02:44:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/1009280", "https://ndownloader.figshare.com/files/1009290", "https://ndownloader.figshare.com/files/1009293", "https://ndownloader.figshare.com/files/1009295", "https://ndownloader.figshare.com/files/1009296", "https://ndownloader.figshare.com/files/1009298"], "description"=>"<div><p>Morphological analysis of mitotic chromosomes is used to detect mutagenic chemical compounds and to estimate the dose of ionizing radiation to be administered. It has long been believed that chromosomal breaks are always associated with double-strand breaks (DSBs). We here provide compelling evidence against this canonical theory. We employed a genetic approach using two cell lines, chicken DT40 and human Nalm-6. We measured the number of chromosomal breaks induced by three replication-blocking agents (aphidicolin, 5-fluorouracil, and hydroxyurea) in DSB-repair-proficient <i>wild-type</i> cells and cells deficient in both homologous recombination and nonhomologous end-joining (the two major DSB-repair pathways). Exposure of cells to the three replication-blocking agents for at least two cell cycles resulted in comparable numbers of chromosomal breaks for <i>RAD54<sup>−/−/</sup>KU70<sup>−/−</sup></i> DT40 clones and <i>wild-type</i> cells. Likewise, the numbers of chromosomal breaks induced in <i>RAD54<sup>−/−/</sup>LIG4<sup>−/−</sup></i> Nalm-6 clones and <i>wild-type</i> cells were also comparable. These data indicate that the replication-blocking agents can cause chromosomal breaks unassociated with DSBs. In contrast with DSB-repair-deficient cells, chicken DT40 cells deficient in PIF1 or ATRIP, which molecules contribute to the completion of DNA replication, displayed higher numbers of mitotic chromosomal breaks induced by aphidicolin than did <i>wild-type</i> cells, suggesting that single-strand gaps left unreplicated may result in mitotic chromosomal breaks.</p> </div>", "links"=>[], "tags"=>["dna", "replication", "mitotic", "chromosomal", "breakage", "unassociated", "double-strand"], "article_id"=>669880, "categories"=>["Molecular Biology", "Biochemistry", "Cancer", "Cell Biology", "Genetics"], "users"=>["Mari Fujita", "Hiroyuki Sasanuma", "Kimiyo N. Yamamoto", "Hiroshi Harada", "Aya Kurosawa", "Noritaka Adachi", "Masato Omura", "Masahiro Hiraoka", "Shunichi Takeda", "Kouji Hirota"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0060043.s001", "https://dx.doi.org/10.1371/journal.pone.0060043.s002", "https://dx.doi.org/10.1371/journal.pone.0060043.s003", "https://dx.doi.org/10.1371/journal.pone.0060043.s004", "https://dx.doi.org/10.1371/journal.pone.0060043.s005", "https://dx.doi.org/10.1371/journal.pone.0060043.s006"], "stats"=>{"downloads"=>2, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Interference_in_DNA_Replication_Can_Cause_Mitotic_Chromosomal_Breakage_Unassociated_with_Double_Strand_Breaks_/669880", "title"=>"Interference in DNA Replication Can Cause Mitotic Chromosomal Breakage Unassociated with Double-Strand Breaks", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-04-03 02:44:40"}

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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/Agriculture", "average_usage"=>[241, 422, 551, 659, 765, 866, 954, 1043, 1142, 1235, 1311, 1397, 1462]}, {"subject_area"=>"/Biology and life sciences/Organisms", "average_usage"=>[281, 484, 611, 728, 835, 934, 1030, 1123, 1214, 1299, 1383, 1464]}, {"subject_area"=>"/Physical sciences/Physics", "average_usage"=>[254, 421, 527, 626, 720, 813, 900, 983, 1063, 1136, 1210, 1283, 1342]}]}
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