A Major Role of the RecFOR Pathway in DNA Double-Strand-Break Repair through ESDSA in Deinococcus radiodurans
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{"title"=>"A major role of the RecFOR pathway in DNA double-strand-break repair through ESDSA in Deinococcus radiodurans", "type"=>"journal", "authors"=>[{"first_name"=>"Esma", "last_name"=>"Bentchikou", "scopus_author_id"=>"16743486500"}, {"first_name"=>"Pascale", "last_name"=>"Servant", "scopus_author_id"=>"6603921161"}, {"first_name"=>"Genevie", "last_name"=>"Coste", "scopus_author_id"=>"6602090231"}, {"first_name"=>"Suzanne", "last_name"=>"Sommer", "scopus_author_id"=>"7201437144"}], "year"=>2010, "source"=>"PLoS Genetics", "identifiers"=>{"pui"=>"358308123", "sgr"=>"76749109918", "pmid"=>"20090937", "scopus"=>"2-s2.0-76749109918", "isbn"=>"1553-7404 (Electronic)\\n1553-7390 (Linking)", "doi"=>"10.1371/journal.pgen.1000774", "issn"=>"15537390"}, "id"=>"9afa596a-7545-3ebe-acd0-4d873a96dfa8", "abstract"=>"In Deinococcus radiodurans, the extreme resistance to DNA-shattering treatments such as ionizing radiation or desiccation is correlated with its ability to reconstruct a functional genome from hundreds of chromosomal fragments. The rapid reconstitution of an intact genome is thought to occur through an extended synthesis-dependent strand annealing process (ESDSA) followed by DNA recombination. Here, we investigated the role of key components of the RecF pathway in ESDSA in this organism naturally devoid of RecB and RecC proteins. We demonstrate that inactivation of RecJ exonuclease results in cell lethality, indicating that this protein plays a key role in genome maintenance. Cells devoid of RecF, RecO, or RecR proteins also display greatly impaired growth and an important lethal sectoring as bacteria devoid of RecA protein. Other aspects of the phenotype of recFOR knock-out mutants paralleled that of a DeltarecA mutant: DeltarecFOR mutants are extremely radiosensitive and show a slow assembly of radiation-induced chromosomal fragments, not accompanied by DNA synthesis, and reduced DNA degradation. Cells devoid of RecQ, the major helicase implicated in repair through the RecF pathway in E. coli, are resistant to gamma-irradiation and have a wild-type DNA repair capacity as also shown for cells devoid of the RecD helicase; in contrast, DeltauvrD mutants show a markedly decreased radioresistance, an increased latent period in the kinetics of DNA double-strand-break repair, and a slow rate of fragment assembly correlated with a slow rate of DNA synthesis. Combining RecQ or RecD deficiency with UvrD deficiency did not significantly accentuate the phenotype of DeltauvrD mutants. In conclusion, RecFOR proteins are essential for DNA double-strand-break repair through ESDSA whereas RecJ protein is essential for cell viability and UvrD helicase might be involved in the processing of double stranded DNA ends and/or in the DNA synthesis step of ESDSA.", "link"=>"http://www.mendeley.com/research/major-role-recfor-pathway-dna-doublestrandbreak-repair-through-esdsa-deinococcus-radiodurans", "reader_count"=>89, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>4, "Researcher"=>19, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>24, "Student > Postgraduate"=>4, "Other"=>3, "Student > Master"=>15, "Student > Bachelor"=>11, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>4, "Researcher"=>19, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>24, "Student > Postgraduate"=>4, "Other"=>3, "Student > Master"=>15, "Student > Bachelor"=>11, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>5, "Engineering"=>1, "Environmental Science"=>3, "Biochemistry, Genetics and Molecular Biology"=>13, "Agricultural and Biological Sciences"=>61, "Medicine and Dentistry"=>1, "Neuroscience"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Physics and Astronomy"=>2, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Neuroscience"=>{"Neuroscience"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>61}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>13}, "Unspecified"=>{"Unspecified"=>5}, "Environmental Science"=>{"Environmental Science"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"United States"=>2, "Japan"=>1, "France"=>1, "Chile"=>1, "Tunisia"=>1, "Portugal"=>1, "India"=>2}, "group_count"=>2}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/867561"], "description"=>"<p>(A) Kinetics of DSB repair in wild-type, Δ<i>recA</i>, Δ<i>recF</i>, Δ<i>recO</i>, and Δ<i>recR</i> mutants. PFGE shows <i>NotI</i> treated DNA from unirradiated cells (lane pre-irradiation) and from irradiated cells (6,8kGy) immediately after irradiation (0) and at the indicated incubation times (hours). (B) Rate of DNA synthesis in wild-type, Δ<i>recA</i>, Δ<i>recF</i>, Δ<i>recO</i>, and Δ<i>recR</i> mutants. Incorporation of [<sup>3</sup>H]thymidine during 15-min pulse labelling measures the global rate of DNA synthesis in irradiated (filled circles) and unirradiated (circles) bacteria.</p>", "links"=>[], "tags"=>["dna", "synthesis"], "article_id"=>538020, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1000774.g006", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DNA_repair_and_DNA_synthesis_in_recA_recF_recO_and_recR_mutants_/538020", "title"=>"DNA repair and DNA synthesis in Δ<i>recA</i>, Δ<i>recF</i>, Δ<i>recO</i>, and Δ<i>recR</i> mutants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-01-15 02:13:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/867280"], "description"=>"<p>Bacterial strains GY12965 (Δ<i>recF</i>, open circles), GY 12966 (Δ<i>recO</i>, open squares), GY 12967 (Δ<i>recR</i>, filled triangles), GY 12968 (Δ<i>recA</i>, filled circles), and wild-type (closed squares) were incubated at 30°C. At different times, incubation samples were taken and the A<sub>650</sub> values of the cultures and the numbers of viable cells/ml were measured.</p>", "links"=>[], "tags"=>["cells", "devoid", "genes", "reduced", "plating"], "article_id"=>537738, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1000774.g003", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_D_radiodurans_cells_devoid_of_recA_recF_recO_or_recR_genes_show_reduced_plating_efficiency_/537738", "title"=>"<i>D. radiodurans</i> cells devoid of <i>recA</i>, <i>recF</i>, <i>recO</i>, or <i>recR</i> genes show reduced plating efficiency.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-01-15 02:08:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/867411"], "description"=>"<p>(A) Kinetics of DSB repair in wild-type, Δ<i>recQ</i>, Δ<i>recD</i>, and Δ<i>uvrD</i> mutants followed by pulsed-field gel electrophoresis (PFGE). PFGE shows <i>NotI</i> treated DNA from unirradiated cells (lane pre-irradiation) and from irradiated cells (6,8 kGy) immediately after irradiation (0) and at the indicated incubation times (hours). (B) Rate of DNA synthesis in wild-type, Δ<i>recQ</i>, Δ<i>recD</i>, and Δ<i>uvrD</i> mutants. Incorporation of [<sup>3</sup>H]thymidine during 15-min pulse labelling measures the global rate of DNA synthesis in 6.8 kGy-irradiated (filled circles) and unirradiated (open circles) bacteria.</p>", "links"=>[], "tags"=>["dna", "synthesis"], "article_id"=>537871, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1000774.g005", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DNA_repair_and_DNA_synthesis_in_recQ_recD_and_uvrD_mutants_/537871", "title"=>"DNA repair and DNA synthesis in Δ<i>recQ</i>, Δ<i>recD</i>, and Δ<i>uvrD</i> mutants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-01-15 02:11:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/867773"], "description"=>"<p>Plasmids.</p>", "links"=>[], "tags"=>["cell biology/cellular death and stress responses", "genetics and genomics/gene function", "microbiology/microbial growth and development", "molecular biology/dna repair", "molecular biology/dna replication", "molecular biology/recombination"], "article_id"=>538225, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1000774.t002", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Plasmids_/538225", "title"=>"Plasmids.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2010-01-15 02:17:05"}
  • {"files"=>["https://ndownloader.figshare.com/files/867812"], "description"=>"<p>Bacterial strains.</p>", "links"=>[], "tags"=>["cell biology/cellular death and stress responses", "genetics and genomics/gene function", "microbiology/microbial growth and development", "molecular biology/dna repair", "molecular biology/dna replication", "molecular biology/recombination"], "article_id"=>538265, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1000774.t001", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bacterial_strains_/538265", "title"=>"Bacterial strains.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2010-01-15 02:17:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/867168"], "description"=>"<p>Strains were grown in liquid medium with spectinomycin at 28°C. The dilutions of cells were spotted on medium with or without spectinomycin at 28°C (A) or 37°C (B). Lane 1–3: strain GY14110 [Δ<i>recJ</i> (p<i>repU</i>Ts-<i>recJ<sup>+</sup></i>)], lane 4: strain GY13781 containing thermosensitive plasmid p13840 (p<i>repU</i>Ts), lane 5: strain GY13786 containing non-thermosensitive plasmid p11554 (p<i>repU</i>).</p>", "links"=>[], "tags"=>["cell biology/cellular death and stress responses", "genetics and genomics/gene function", "microbiology/microbial growth and development", "molecular biology/dna repair", "molecular biology/dna replication", "molecular biology/recombination"], "article_id"=>537624, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1000774.g002", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_recJ_is_essential_for_D_radiodurans_viability_/537624", "title"=>"<i>recJ</i> is essential for <i>D. radiodurans</i> viability.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-01-15 02:07:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/867104"], "description"=>"<p>(A) Schematic representation of the allele replacement event in <i>recJ</i> gene. Short arrows indicate the position of specific primers used for diagnostic PCR. Primers are described in <a href=\"http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1000774#pgen.1000774.s004\" target=\"_blank\">Table S1</a>. (B) PCR analysis of three independent candidate <i>recJ</i> mutants and three independent Δ<i>recJ</i> (p<i>repU</i>TS-<i>recJ</i><sup>+</sup>) mutants.</p>", "links"=>[], "tags"=>["deletion-substitution", "radiodurans"], "article_id"=>537561, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1000774.g001", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_representation_and_test_of_deletion_substitution_in_the_D_radiodurans_recJ_gene_/537561", "title"=>"Schematic representation and test of deletion-substitution in the <i>D. radiodurans recJ</i> gene.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-01-15 02:06:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/867328"], "description"=>"<p>(A) Increased sensitivity of cells devoid of <i>uvrD</i> gene. R1 (wild-type, open inverted triangles), GY12957 (Δ<i>recQ</i>, filled squares), GY12974 (Δ<i>uvrD</i>, filled circles), GY12975 (Δ<i>recQ</i>Δ<i>uvrD</i>, open squares), GY12976 (Δ<i>recD</i>Δ<i>uvrD</i>, filled diamonds), GY12977 (Δ<i>recQ</i>Δ<i>recD</i>, filled triangles), GY13130 (Δ<i>recD</i>, open circles) bacteria were exposed to γ-irradiation at doses indicated on the abscissa; and cell survival was measured as described in the <a href=\"http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1000774#s4\" target=\"_blank\">Materials and Methods</a>. (B) Δ<i>recFOR</i> mutants are as sensitive as Δ<i>recA</i> mutant to γ-irradiation. Bacterial strains GY12936 (wild-type/p11520, inverted triangles), GY14115 (Δ<i>recA</i>/p11559, filled circles), GY14116 (Δ<i>recO</i>/p11520, filled diamonds), GY14117 (Δ<i>recF</i>/p11520, filled squares), GY14118 (Δ<i>recR</i>/p11520, filled triangles), GY14111 (Δ<i>recA</i>/p11562: <i>recA</i><sup>+</sup>, open circles), GY14112 (Δ<i>recO</i>/p11860: <i>recO</i><sup>+</sup>, open diamonds), GY14113 (Δ<i>recF</i>/p11862: <i>recF</i><sup>+</sup>, open squares), GY14114 (Δ<i>recR</i>/p11870: <i>recR</i><sup>+</sup>, open triangles) were exposed to γ-irradiation at doses indicated on the abscissa, and cell survival was measured as described in the <a href=\"http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1000774#s4\" target=\"_blank\">Materials and Methods</a>.</p>", "links"=>[], "tags"=>["cells", "devoid"], "article_id"=>537786, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1000774.g004", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Increased_sensitivity_to_947_irradiation_of_cells_devoid_of_RecF_RecO_RecR_or_UvrD_/537786", "title"=>"Increased sensitivity to γ-irradiation of cells devoid of RecF, RecO, RecR, or UvrD.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-01-15 02:09:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/867701"], "description"=>"<p>Model of initiation of DNA double-strand-break repair through ESDSA in <i>D. radiodurans</i>.</p>", "links"=>[], "tags"=>["initiation", "dna", "double-strand-break", "esdsa"], "article_id"=>538159, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>"https://dx.doi.org/10.1371/journal.pgen.1000774.g007", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Model_of_initiation_of_DNA_double_strand_break_repair_through_ESDSA_in_D_radiodurans_/538159", "title"=>"Model of initiation of DNA double-strand-break repair through ESDSA in <i>D. radiodurans</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-01-15 02:15:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/430663", "https://ndownloader.figshare.com/files/430690", "https://ndownloader.figshare.com/files/430743", "https://ndownloader.figshare.com/files/430765"], "description"=>"<div><p>In <em>Deinococcus radiodurans</em>, the extreme resistance to DNA–shattering treatments such as ionizing radiation or desiccation is correlated with its ability to reconstruct a functional genome from hundreds of chromosomal fragments. The rapid reconstitution of an intact genome is thought to occur through an extended synthesis-dependent strand annealing process (ESDSA) followed by DNA recombination. Here, we investigated the role of key components of the RecF pathway in ESDSA in this organism naturally devoid of RecB and RecC proteins. We demonstrate that inactivation of RecJ exonuclease results in cell lethality, indicating that this protein plays a key role in genome maintenance. Cells devoid of RecF, RecO, or RecR proteins also display greatly impaired growth and an important lethal sectoring as bacteria devoid of RecA protein. Other aspects of the phenotype of <em>recFOR</em> knock-out mutants paralleled that of a Δ<em>recA</em> mutant: Δ<em>recFOR</em> mutants are extremely radiosensitive and show a slow assembly of radiation-induced chromosomal fragments, not accompanied by DNA synthesis, and reduced DNA degradation. Cells devoid of RecQ, the major helicase implicated in repair through the RecF pathway in <em>E. coli</em>, are resistant to γ-irradiation and have a wild-type DNA repair capacity as also shown for cells devoid of the RecD helicase; in contrast, Δ<em>uvrD</em> mutants show a markedly decreased radioresistance, an increased latent period in the kinetics of DNA double-strand-break repair, and a slow rate of fragment assembly correlated with a slow rate of DNA synthesis. Combining RecQ or RecD deficiency with UvrD deficiency did not significantly accentuate the phenotype of Δ<em>uvrD</em> mutants. In conclusion, RecFOR proteins are essential for DNA double-strand-break repair through ESDSA whereas RecJ protein is essential for cell viability and UvrD helicase might be involved in the processing of double stranded DNA ends and/or in the DNA synthesis step of ESDSA.</p></div>", "links"=>[], "tags"=>["recfor", "pathway", "dna", "double-strand-break", "esdsa"], "article_id"=>144990, "categories"=>["Molecular Biology", "Cell Biology", "Genetics", "Microbiology"], "users"=>["Esma Bentchikou", "Pascale Servant", "Geneviève Coste", "Suzanne Sommer"], "doi"=>["https://dx.doi.org/10.1371/journal.pgen.1000774.s001", "https://dx.doi.org/10.1371/journal.pgen.1000774.s002", "https://dx.doi.org/10.1371/journal.pgen.1000774.s003", "https://dx.doi.org/10.1371/journal.pgen.1000774.s004"], "stats"=>{"downloads"=>21, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/A_Major_Role_of_the_RecFOR_Pathway_in_DNA_Double_Strand_Break_Repair_through_ESDSA_in_Deinococcus_radiodurans_/144990", "title"=>"A Major Role of the RecFOR Pathway in DNA Double-Strand-Break Repair through ESDSA in <em>Deinococcus radiodurans</em>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2010-01-15 01:23:10"}

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  • {"unique-ip"=>"8", "full-text"=>"8", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"6"}
  • {"unique-ip"=>"14", "full-text"=>"10", "pdf"=>"6", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2018", "month"=>"7"}
  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"8"}
  • {"unique-ip"=>"3", "full-text"=>"2", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
  • {"unique-ip"=>"14", "full-text"=>"14", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
  • {"unique-ip"=>"13", "full-text"=>"12", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
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  • {"unique-ip"=>"7", "full-text"=>"6", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
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Relative Metric

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