DNA end resection by Dna2–Sgs1–RPA and its stimulation by Top3–Rmi1 and Mre11–Rad50–Xrs2 View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2010-09

AUTHORS

Petr Cejka, Elda Cannavo, Piotr Polaczek, Taro Masuda-Sasa, Subhash Pokharel, Judith L. Campbell, Stephen C. Kowalczykowski

ABSTRACT

The repair of DNA double-strand breaks (DSBs) by homologous recombination requires processing of broken ends. For repair to start, the DSB must first be resected to generate a 3'-single-stranded DNA (ssDNA) overhang, which becomes a substrate for the DNA strand exchange protein, Rad51 (ref. 1). Genetic studies have implicated a multitude of proteins in the process, including helicases, nucleases and topoisomerases. Here we biochemically reconstitute elements of the resection process and reveal that it requires the nuclease Dna2, the RecQ-family helicase Sgs1 and the ssDNA-binding protein replication protein-A (RPA). We establish that Dna2, Sgs1 and RPA constitute a minimal protein complex capable of DNA resection in vitro. Sgs1 helicase unwinds the DNA to produce an intermediate that is digested by Dna2, and RPA stimulates DNA unwinding by Sgs1 in a species-specific manner. Interestingly, RPA is also required both to direct Dna2 nucleolytic activity to the 5'-terminated strand of the DNA break and to inhibit 3' to 5' degradation by Dna2, actions that generate and protect the 3'-ssDNA overhang, respectively. In addition to this core machinery, we establish that both the topoisomerase 3 (Top3) and Rmi1 complex and the Mre11-Rad50-Xrs2 complex (MRX) have important roles as stimulatory components. Stimulation of end resection by the Top3-Rmi1 heterodimer and the MRX proteins is by complex formation with Sgs1 (refs 5, 6), which unexpectedly stimulates DNA unwinding. We suggest that Top3-Rmi1 and MRX are important for recruitment of the Sgs1-Dna2 complex to DSBs. Our experiments provide a mechanistic framework for understanding the initial steps of recombinational DNA repair in eukaryotes. More... »

PAGES

112

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nature09355

DOI

http://dx.doi.org/10.1038/nature09355

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1049775672

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/20811461


Indexing Status Check whether this publication has been indexed by Scopus and Web Of Science using the SN Indexing Status Tool
Incoming Citations Browse incoming citations for this publication using opencitations.net

JSON-LD is the canonical representation for SciGraph data.

TIP: You can open this SciGraph record using an external JSON-LD service: JSON-LD Playground Google SDTT

[
  {
    "@context": "https://springernature.github.io/scigraph/jsonld/sgcontext.json", 
    "about": [
      {
        "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/0601", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Biochemistry and Cell Biology", 
        "type": "DefinedTerm"
      }, 
      {
        "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/06", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Biological Sciences", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "DNA Helicases", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "DNA Repair", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "DNA-Binding Proteins", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Deoxyribonucleases", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Endodeoxyribonucleases", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Exodeoxyribonucleases", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "RecQ Helicases", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Replication Protein A", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Saccharomyces cerevisiae", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Saccharomyces cerevisiae Proteins", 
        "type": "DefinedTerm"
      }
    ], 
    "author": [
      {
        "affiliation": {
          "alternateName": "University of California, Davis", 
          "id": "https://www.grid.ac/institutes/grid.27860.3b", 
          "name": [
            "Department of Microbiology, University of California, Davis, Davis, California 95616-8665, USA", 
            "Department of Molecular and Cellular Biology, University of California, Davis, Davis, California 95616-8665, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Cejka", 
        "givenName": "Petr", 
        "id": "sg:person.01364612220.22", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01364612220.22"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "University of California, Davis", 
          "id": "https://www.grid.ac/institutes/grid.27860.3b", 
          "name": [
            "Department of Microbiology, University of California, Davis, Davis, California 95616-8665, USA", 
            "Department of Molecular and Cellular Biology, University of California, Davis, Davis, California 95616-8665, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Cannavo", 
        "givenName": "Elda", 
        "id": "sg:person.0776606706.81", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0776606706.81"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "California Institute of Technology", 
          "id": "https://www.grid.ac/institutes/grid.20861.3d", 
          "name": [
            "Division of Biology, California Institute of Technology, Pasadena, California 91125, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Polaczek", 
        "givenName": "Piotr", 
        "id": "sg:person.01304354241.53", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01304354241.53"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "California Institute of Technology", 
          "id": "https://www.grid.ac/institutes/grid.20861.3d", 
          "name": [
            "Division of Biology, California Institute of Technology, Pasadena, California 91125, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Masuda-Sasa", 
        "givenName": "Taro", 
        "id": "sg:person.01216102435.96", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01216102435.96"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "California Institute of Technology", 
          "id": "https://www.grid.ac/institutes/grid.20861.3d", 
          "name": [
            "Division of Biology, California Institute of Technology, Pasadena, California 91125, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Pokharel", 
        "givenName": "Subhash", 
        "id": "sg:person.01022044161.65", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01022044161.65"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "California Institute of Technology", 
          "id": "https://www.grid.ac/institutes/grid.20861.3d", 
          "name": [
            "Division of Biology, California Institute of Technology, Pasadena, California 91125, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Campbell", 
        "givenName": "Judith L.", 
        "id": "sg:person.0642141210.65", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0642141210.65"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "University of California, Davis", 
          "id": "https://www.grid.ac/institutes/grid.27860.3b", 
          "name": [
            "Department of Microbiology, University of California, Davis, Davis, California 95616-8665, USA", 
            "Department of Molecular and Cellular Biology, University of California, Davis, Davis, California 95616-8665, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Kowalczykowski", 
        "givenName": "Stephen C.", 
        "id": "sg:person.01372345264.09", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01372345264.09"
        ], 
        "type": "Person"
      }
    ], 
    "citation": [
      {
        "id": "sg:pub.10.1038/nsmb.1919", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1002138390", 
          "https://doi.org/10.1038/nsmb.1919"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nsmb.1919", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1002138390", 
          "https://doi.org/10.1038/nsmb.1919"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1128/mcb.25.13.5738-5751.2005", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1004857820"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1101/gad.12.8.1134", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1007603368"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1038/sj.emboj.7600684", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1008403264"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1038/sj.emboj.7600684", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1008403264"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1074/jbc.m802244200", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1009204427"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1128/mcb.24.21.9682-9694.2004", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1010422397"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1074/jbc.270.45.26766", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1012451568"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1073/pnas.0809380105", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1014592014"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1093/emboj/cdg391", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1016529319"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1101/gad.503108", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1017396089"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1006/jmbi.2001.4555", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1018601565"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1074/jbc.m313216200", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1020867535"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nature07312", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1021228402", 
          "https://doi.org/10.1038/nature07312"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.molcel.2008.11.021", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1023368902"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1128/mcb.14.12.8391", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1024726124"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nature09318", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1027803050", 
          "https://doi.org/10.1038/nature09318"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nature09318", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1027803050", 
          "https://doi.org/10.1038/nature09318"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1146/annurev.genet.38.072902.091500", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1028767256"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1128/mcb.25.11.4476-4487.2005", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1029059749"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.cell.2008.08.037", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1029629280"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1074/jbc.m909511199", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1029961439"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1074/jbc.m109.083196", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1030056442"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.dnarep.2004.03.014", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1030622956"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nature03872", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1031191047", 
          "https://doi.org/10.1038/nature03872"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nature03872", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1031191047", 
          "https://doi.org/10.1038/nature03872"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/0022-2836(92)90681-9", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1032181633"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1101/gad.1780709", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1033256124"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/s0092-8674(00)80315-3", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1034604776"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1038/emboj.2008.111", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1036540030"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1074/jbc.m009471200", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1037454010"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1128/mmbr.00020-08", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1041476570"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1371/journal.pone.0004267", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1041896408"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1093/nar/gkg422", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1044580340"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.molcel.2009.09.036", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1047783751"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1074/jbc.m006513200", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1048933642"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/35086609", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1049047000", 
          "https://doi.org/10.1038/35086609"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/35086609", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1049047000", 
          "https://doi.org/10.1038/35086609"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1126/science.7855597", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1062649921"
        ], 
        "type": "CreativeWork"
      }
    ], 
    "datePublished": "2010-09", 
    "datePublishedReg": "2010-09-01", 
    "description": "The repair of DNA double-strand breaks (DSBs) by homologous recombination requires processing of broken ends. For repair to start, the DSB must first be resected to generate a 3'-single-stranded DNA (ssDNA) overhang, which becomes a substrate for the DNA strand exchange protein, Rad51 (ref. 1). Genetic studies have implicated a multitude of proteins in the process, including helicases, nucleases and topoisomerases. Here we biochemically reconstitute elements of the resection process and reveal that it requires the nuclease Dna2, the RecQ-family helicase Sgs1 and the ssDNA-binding protein replication protein-A (RPA). We establish that Dna2, Sgs1 and RPA constitute a minimal protein complex capable of DNA resection in vitro. Sgs1 helicase unwinds the DNA to produce an intermediate that is digested by Dna2, and RPA stimulates DNA unwinding by Sgs1 in a species-specific manner. Interestingly, RPA is also required both to direct Dna2 nucleolytic activity to the 5'-terminated strand of the DNA break and to inhibit 3' to 5' degradation by Dna2, actions that generate and protect the 3'-ssDNA overhang, respectively. In addition to this core machinery, we establish that both the topoisomerase 3 (Top3) and Rmi1 complex and the Mre11-Rad50-Xrs2 complex (MRX) have important roles as stimulatory components. Stimulation of end resection by the Top3-Rmi1 heterodimer and the MRX proteins is by complex formation with Sgs1 (refs 5, 6), which unexpectedly stimulates DNA unwinding. We suggest that Top3-Rmi1 and MRX are important for recruitment of the Sgs1-Dna2 complex to DSBs. Our experiments provide a mechanistic framework for understanding the initial steps of recombinational DNA repair in eukaryotes.", 
    "genre": "research_article", 
    "id": "sg:pub.10.1038/nature09355", 
    "inLanguage": [
      "en"
    ], 
    "isAccessibleForFree": true, 
    "isFundedItemOf": [
      {
        "id": "sg:grant.2512730", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.2519325", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.2516920", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.2635378", 
        "type": "MonetaryGrant"
      }
    ], 
    "isPartOf": [
      {
        "id": "sg:journal.1018957", 
        "issn": [
          "0090-0028", 
          "1476-4687"
        ], 
        "name": "Nature", 
        "type": "Periodical"
      }, 
      {
        "issueNumber": "7311", 
        "type": "PublicationIssue"
      }, 
      {
        "type": "PublicationVolume", 
        "volumeNumber": "467"
      }
    ], 
    "name": "DNA end resection by Dna2\u2013Sgs1\u2013RPA and its stimulation by Top3\u2013Rmi1 and Mre11\u2013Rad50\u2013Xrs2", 
    "pagination": "112", 
    "productId": [
      {
        "name": "readcube_id", 
        "type": "PropertyValue", 
        "value": [
          "ed1dd35f76ed1ffb730a3a6dd12f67a85931f8dc29dbc4383163dc2402a3705e"
        ]
      }, 
      {
        "name": "pubmed_id", 
        "type": "PropertyValue", 
        "value": [
          "20811461"
        ]
      }, 
      {
        "name": "nlm_unique_id", 
        "type": "PropertyValue", 
        "value": [
          "0410462"
        ]
      }, 
      {
        "name": "doi", 
        "type": "PropertyValue", 
        "value": [
          "10.1038/nature09355"
        ]
      }, 
      {
        "name": "dimensions_id", 
        "type": "PropertyValue", 
        "value": [
          "pub.1049775672"
        ]
      }
    ], 
    "sameAs": [
      "https://doi.org/10.1038/nature09355", 
      "https://app.dimensions.ai/details/publication/pub.1049775672"
    ], 
    "sdDataset": "articles", 
    "sdDatePublished": "2019-04-11T10:50", 
    "sdLicense": "https://scigraph.springernature.com/explorer/license/", 
    "sdPublisher": {
      "name": "Springer Nature - SN SciGraph project", 
      "type": "Organization"
    }, 
    "sdSource": "s3://com-uberresearch-data-dimensions-target-20181106-alternative/cleanup/v134/2549eaecd7973599484d7c17b260dba0a4ecb94b/merge/v9/a6c9fde33151104705d4d7ff012ea9563521a3ce/jats-lookup/v90/0000000350_0000000350/records_77577_00000000.jsonl", 
    "type": "ScholarlyArticle", 
    "url": "https://www.nature.com/articles/nature09355"
  }
]
 

Download the RDF metadata as:  json-ld nt turtle xml License info

HOW TO GET THIS DATA PROGRAMMATICALLY:

JSON-LD is a popular format for linked data which is fully compatible with JSON.

curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1038/nature09355'

N-Triples is a line-based linked data format ideal for batch operations.

curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1038/nature09355'

Turtle is a human-readable linked data format.

curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1038/nature09355'

RDF/XML is a standard XML format for linked data.

curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1038/nature09355'


 

This table displays all metadata directly associated to this object as RDF triples.

273 TRIPLES      21 PREDICATES      74 URIs      31 LITERALS      19 BLANK NODES

Subject Predicate Object
1 sg:pub.10.1038/nature09355 schema:about N0bc03caabfc547b6971aa2c0812d0d21
2 N3990c1f7575f42c1a1cdaf59c566ef12
3 N67f9fceaf46b4e40a78ff4b13a322185
4 N7b21a0262d3f4c55968b284aeec9bd19
5 Na93321d04c16476bbf3e2eccd123d708
6 Ncec75730816047d1bfb47e7497371e30
7 Nda1052157a56497d89cb42ba4d759d12
8 Ne65937adf63f4026a6838e7acea4e1c1
9 Neaf6d8a57fb8431981bde175d6ada5e8
10 Nfab02d6b76534365a3424ad534922c4d
11 anzsrc-for:06
12 anzsrc-for:0601
13 schema:author N87ef89b164904444bd3f9b8a05f2fde0
14 schema:citation sg:pub.10.1038/35086609
15 sg:pub.10.1038/nature03872
16 sg:pub.10.1038/nature07312
17 sg:pub.10.1038/nature09318
18 sg:pub.10.1038/nsmb.1919
19 https://doi.org/10.1006/jmbi.2001.4555
20 https://doi.org/10.1016/0022-2836(92)90681-9
21 https://doi.org/10.1016/j.cell.2008.08.037
22 https://doi.org/10.1016/j.dnarep.2004.03.014
23 https://doi.org/10.1016/j.molcel.2008.11.021
24 https://doi.org/10.1016/j.molcel.2009.09.036
25 https://doi.org/10.1016/s0092-8674(00)80315-3
26 https://doi.org/10.1038/emboj.2008.111
27 https://doi.org/10.1038/sj.emboj.7600684
28 https://doi.org/10.1073/pnas.0809380105
29 https://doi.org/10.1074/jbc.270.45.26766
30 https://doi.org/10.1074/jbc.m006513200
31 https://doi.org/10.1074/jbc.m009471200
32 https://doi.org/10.1074/jbc.m109.083196
33 https://doi.org/10.1074/jbc.m313216200
34 https://doi.org/10.1074/jbc.m802244200
35 https://doi.org/10.1074/jbc.m909511199
36 https://doi.org/10.1093/emboj/cdg391
37 https://doi.org/10.1093/nar/gkg422
38 https://doi.org/10.1101/gad.12.8.1134
39 https://doi.org/10.1101/gad.1780709
40 https://doi.org/10.1101/gad.503108
41 https://doi.org/10.1126/science.7855597
42 https://doi.org/10.1128/mcb.14.12.8391
43 https://doi.org/10.1128/mcb.24.21.9682-9694.2004
44 https://doi.org/10.1128/mcb.25.11.4476-4487.2005
45 https://doi.org/10.1128/mcb.25.13.5738-5751.2005
46 https://doi.org/10.1128/mmbr.00020-08
47 https://doi.org/10.1146/annurev.genet.38.072902.091500
48 https://doi.org/10.1371/journal.pone.0004267
49 schema:datePublished 2010-09
50 schema:datePublishedReg 2010-09-01
51 schema:description The repair of DNA double-strand breaks (DSBs) by homologous recombination requires processing of broken ends. For repair to start, the DSB must first be resected to generate a 3'-single-stranded DNA (ssDNA) overhang, which becomes a substrate for the DNA strand exchange protein, Rad51 (ref. 1). Genetic studies have implicated a multitude of proteins in the process, including helicases, nucleases and topoisomerases. Here we biochemically reconstitute elements of the resection process and reveal that it requires the nuclease Dna2, the RecQ-family helicase Sgs1 and the ssDNA-binding protein replication protein-A (RPA). We establish that Dna2, Sgs1 and RPA constitute a minimal protein complex capable of DNA resection in vitro. Sgs1 helicase unwinds the DNA to produce an intermediate that is digested by Dna2, and RPA stimulates DNA unwinding by Sgs1 in a species-specific manner. Interestingly, RPA is also required both to direct Dna2 nucleolytic activity to the 5'-terminated strand of the DNA break and to inhibit 3' to 5' degradation by Dna2, actions that generate and protect the 3'-ssDNA overhang, respectively. In addition to this core machinery, we establish that both the topoisomerase 3 (Top3) and Rmi1 complex and the Mre11-Rad50-Xrs2 complex (MRX) have important roles as stimulatory components. Stimulation of end resection by the Top3-Rmi1 heterodimer and the MRX proteins is by complex formation with Sgs1 (refs 5, 6), which unexpectedly stimulates DNA unwinding. We suggest that Top3-Rmi1 and MRX are important for recruitment of the Sgs1-Dna2 complex to DSBs. Our experiments provide a mechanistic framework for understanding the initial steps of recombinational DNA repair in eukaryotes.
52 schema:genre research_article
53 schema:inLanguage en
54 schema:isAccessibleForFree true
55 schema:isPartOf N5dfd77e2d01b42118f2b8cdeef0da9f7
56 Nddcf52e00ad14d8294c372af8c89c851
57 sg:journal.1018957
58 schema:name DNA end resection by Dna2–Sgs1–RPA and its stimulation by Top3–Rmi1 and Mre11–Rad50–Xrs2
59 schema:pagination 112
60 schema:productId N47fe2e0a13ad40dba097cb61802538a7
61 N6409b84b76754e76be8f181f4e33cc4f
62 N761c2ff593724a2bb19d87ca6d7b31b8
63 Nb6929e47d4744ba8b6039cc02b7694f7
64 Nddef671c0e16449a9d1dc9ea94fbcb0a
65 schema:sameAs https://app.dimensions.ai/details/publication/pub.1049775672
66 https://doi.org/10.1038/nature09355
67 schema:sdDatePublished 2019-04-11T10:50
68 schema:sdLicense https://scigraph.springernature.com/explorer/license/
69 schema:sdPublisher Nb163b85f4afe4e1f8b02b35ee35507bc
70 schema:url https://www.nature.com/articles/nature09355
71 sgo:license sg:explorer/license/
72 sgo:sdDataset articles
73 rdf:type schema:ScholarlyArticle
74 N05c8ca463f3a4035ac96c03969d6e839 rdf:first sg:person.01022044161.65
75 rdf:rest N1bb47b8f636f4c2894cbba32b515d9a0
76 N0bc03caabfc547b6971aa2c0812d0d21 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
77 schema:name Endodeoxyribonucleases
78 rdf:type schema:DefinedTerm
79 N1bb47b8f636f4c2894cbba32b515d9a0 rdf:first sg:person.0642141210.65
80 rdf:rest N46e061cc60ef4ad0b924efaf267e29c6
81 N3990c1f7575f42c1a1cdaf59c566ef12 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
82 schema:name DNA Helicases
83 rdf:type schema:DefinedTerm
84 N46e061cc60ef4ad0b924efaf267e29c6 rdf:first sg:person.01372345264.09
85 rdf:rest rdf:nil
86 N47fe2e0a13ad40dba097cb61802538a7 schema:name dimensions_id
87 schema:value pub.1049775672
88 rdf:type schema:PropertyValue
89 N5dfd77e2d01b42118f2b8cdeef0da9f7 schema:issueNumber 7311
90 rdf:type schema:PublicationIssue
91 N6409b84b76754e76be8f181f4e33cc4f schema:name nlm_unique_id
92 schema:value 0410462
93 rdf:type schema:PropertyValue
94 N67f9fceaf46b4e40a78ff4b13a322185 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
95 schema:name RecQ Helicases
96 rdf:type schema:DefinedTerm
97 N761c2ff593724a2bb19d87ca6d7b31b8 schema:name doi
98 schema:value 10.1038/nature09355
99 rdf:type schema:PropertyValue
100 N7b21a0262d3f4c55968b284aeec9bd19 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
101 schema:name DNA-Binding Proteins
102 rdf:type schema:DefinedTerm
103 N87e28853ba4c4fac9ac3de73726c5455 rdf:first sg:person.01216102435.96
104 rdf:rest N05c8ca463f3a4035ac96c03969d6e839
105 N87ef89b164904444bd3f9b8a05f2fde0 rdf:first sg:person.01364612220.22
106 rdf:rest Nc8cb9086c5434c71baed4fbc8a5d6cb5
107 Na93321d04c16476bbf3e2eccd123d708 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
108 schema:name Deoxyribonucleases
109 rdf:type schema:DefinedTerm
110 Nac85260cf9214412ad412a4dd8c0ae70 rdf:first sg:person.01304354241.53
111 rdf:rest N87e28853ba4c4fac9ac3de73726c5455
112 Nb163b85f4afe4e1f8b02b35ee35507bc schema:name Springer Nature - SN SciGraph project
113 rdf:type schema:Organization
114 Nb6929e47d4744ba8b6039cc02b7694f7 schema:name pubmed_id
115 schema:value 20811461
116 rdf:type schema:PropertyValue
117 Nc8cb9086c5434c71baed4fbc8a5d6cb5 rdf:first sg:person.0776606706.81
118 rdf:rest Nac85260cf9214412ad412a4dd8c0ae70
119 Ncec75730816047d1bfb47e7497371e30 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
120 schema:name Saccharomyces cerevisiae
121 rdf:type schema:DefinedTerm
122 Nda1052157a56497d89cb42ba4d759d12 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
123 schema:name DNA Repair
124 rdf:type schema:DefinedTerm
125 Nddcf52e00ad14d8294c372af8c89c851 schema:volumeNumber 467
126 rdf:type schema:PublicationVolume
127 Nddef671c0e16449a9d1dc9ea94fbcb0a schema:name readcube_id
128 schema:value ed1dd35f76ed1ffb730a3a6dd12f67a85931f8dc29dbc4383163dc2402a3705e
129 rdf:type schema:PropertyValue
130 Ne65937adf63f4026a6838e7acea4e1c1 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
131 schema:name Replication Protein A
132 rdf:type schema:DefinedTerm
133 Neaf6d8a57fb8431981bde175d6ada5e8 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
134 schema:name Saccharomyces cerevisiae Proteins
135 rdf:type schema:DefinedTerm
136 Nfab02d6b76534365a3424ad534922c4d schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
137 schema:name Exodeoxyribonucleases
138 rdf:type schema:DefinedTerm
139 anzsrc-for:06 schema:inDefinedTermSet anzsrc-for:
140 schema:name Biological Sciences
141 rdf:type schema:DefinedTerm
142 anzsrc-for:0601 schema:inDefinedTermSet anzsrc-for:
143 schema:name Biochemistry and Cell Biology
144 rdf:type schema:DefinedTerm
145 sg:grant.2512730 http://pending.schema.org/fundedItem sg:pub.10.1038/nature09355
146 rdf:type schema:MonetaryGrant
147 sg:grant.2516920 http://pending.schema.org/fundedItem sg:pub.10.1038/nature09355
148 rdf:type schema:MonetaryGrant
149 sg:grant.2519325 http://pending.schema.org/fundedItem sg:pub.10.1038/nature09355
150 rdf:type schema:MonetaryGrant
151 sg:grant.2635378 http://pending.schema.org/fundedItem sg:pub.10.1038/nature09355
152 rdf:type schema:MonetaryGrant
153 sg:journal.1018957 schema:issn 0090-0028
154 1476-4687
155 schema:name Nature
156 rdf:type schema:Periodical
157 sg:person.01022044161.65 schema:affiliation https://www.grid.ac/institutes/grid.20861.3d
158 schema:familyName Pokharel
159 schema:givenName Subhash
160 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01022044161.65
161 rdf:type schema:Person
162 sg:person.01216102435.96 schema:affiliation https://www.grid.ac/institutes/grid.20861.3d
163 schema:familyName Masuda-Sasa
164 schema:givenName Taro
165 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01216102435.96
166 rdf:type schema:Person
167 sg:person.01304354241.53 schema:affiliation https://www.grid.ac/institutes/grid.20861.3d
168 schema:familyName Polaczek
169 schema:givenName Piotr
170 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01304354241.53
171 rdf:type schema:Person
172 sg:person.01364612220.22 schema:affiliation https://www.grid.ac/institutes/grid.27860.3b
173 schema:familyName Cejka
174 schema:givenName Petr
175 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01364612220.22
176 rdf:type schema:Person
177 sg:person.01372345264.09 schema:affiliation https://www.grid.ac/institutes/grid.27860.3b
178 schema:familyName Kowalczykowski
179 schema:givenName Stephen C.
180 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01372345264.09
181 rdf:type schema:Person
182 sg:person.0642141210.65 schema:affiliation https://www.grid.ac/institutes/grid.20861.3d
183 schema:familyName Campbell
184 schema:givenName Judith L.
185 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0642141210.65
186 rdf:type schema:Person
187 sg:person.0776606706.81 schema:affiliation https://www.grid.ac/institutes/grid.27860.3b
188 schema:familyName Cannavo
189 schema:givenName Elda
190 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0776606706.81
191 rdf:type schema:Person
192 sg:pub.10.1038/35086609 schema:sameAs https://app.dimensions.ai/details/publication/pub.1049047000
193 https://doi.org/10.1038/35086609
194 rdf:type schema:CreativeWork
195 sg:pub.10.1038/nature03872 schema:sameAs https://app.dimensions.ai/details/publication/pub.1031191047
196 https://doi.org/10.1038/nature03872
197 rdf:type schema:CreativeWork
198 sg:pub.10.1038/nature07312 schema:sameAs https://app.dimensions.ai/details/publication/pub.1021228402
199 https://doi.org/10.1038/nature07312
200 rdf:type schema:CreativeWork
201 sg:pub.10.1038/nature09318 schema:sameAs https://app.dimensions.ai/details/publication/pub.1027803050
202 https://doi.org/10.1038/nature09318
203 rdf:type schema:CreativeWork
204 sg:pub.10.1038/nsmb.1919 schema:sameAs https://app.dimensions.ai/details/publication/pub.1002138390
205 https://doi.org/10.1038/nsmb.1919
206 rdf:type schema:CreativeWork
207 https://doi.org/10.1006/jmbi.2001.4555 schema:sameAs https://app.dimensions.ai/details/publication/pub.1018601565
208 rdf:type schema:CreativeWork
209 https://doi.org/10.1016/0022-2836(92)90681-9 schema:sameAs https://app.dimensions.ai/details/publication/pub.1032181633
210 rdf:type schema:CreativeWork
211 https://doi.org/10.1016/j.cell.2008.08.037 schema:sameAs https://app.dimensions.ai/details/publication/pub.1029629280
212 rdf:type schema:CreativeWork
213 https://doi.org/10.1016/j.dnarep.2004.03.014 schema:sameAs https://app.dimensions.ai/details/publication/pub.1030622956
214 rdf:type schema:CreativeWork
215 https://doi.org/10.1016/j.molcel.2008.11.021 schema:sameAs https://app.dimensions.ai/details/publication/pub.1023368902
216 rdf:type schema:CreativeWork
217 https://doi.org/10.1016/j.molcel.2009.09.036 schema:sameAs https://app.dimensions.ai/details/publication/pub.1047783751
218 rdf:type schema:CreativeWork
219 https://doi.org/10.1016/s0092-8674(00)80315-3 schema:sameAs https://app.dimensions.ai/details/publication/pub.1034604776
220 rdf:type schema:CreativeWork
221 https://doi.org/10.1038/emboj.2008.111 schema:sameAs https://app.dimensions.ai/details/publication/pub.1036540030
222 rdf:type schema:CreativeWork
223 https://doi.org/10.1038/sj.emboj.7600684 schema:sameAs https://app.dimensions.ai/details/publication/pub.1008403264
224 rdf:type schema:CreativeWork
225 https://doi.org/10.1073/pnas.0809380105 schema:sameAs https://app.dimensions.ai/details/publication/pub.1014592014
226 rdf:type schema:CreativeWork
227 https://doi.org/10.1074/jbc.270.45.26766 schema:sameAs https://app.dimensions.ai/details/publication/pub.1012451568
228 rdf:type schema:CreativeWork
229 https://doi.org/10.1074/jbc.m006513200 schema:sameAs https://app.dimensions.ai/details/publication/pub.1048933642
230 rdf:type schema:CreativeWork
231 https://doi.org/10.1074/jbc.m009471200 schema:sameAs https://app.dimensions.ai/details/publication/pub.1037454010
232 rdf:type schema:CreativeWork
233 https://doi.org/10.1074/jbc.m109.083196 schema:sameAs https://app.dimensions.ai/details/publication/pub.1030056442
234 rdf:type schema:CreativeWork
235 https://doi.org/10.1074/jbc.m313216200 schema:sameAs https://app.dimensions.ai/details/publication/pub.1020867535
236 rdf:type schema:CreativeWork
237 https://doi.org/10.1074/jbc.m802244200 schema:sameAs https://app.dimensions.ai/details/publication/pub.1009204427
238 rdf:type schema:CreativeWork
239 https://doi.org/10.1074/jbc.m909511199 schema:sameAs https://app.dimensions.ai/details/publication/pub.1029961439
240 rdf:type schema:CreativeWork
241 https://doi.org/10.1093/emboj/cdg391 schema:sameAs https://app.dimensions.ai/details/publication/pub.1016529319
242 rdf:type schema:CreativeWork
243 https://doi.org/10.1093/nar/gkg422 schema:sameAs https://app.dimensions.ai/details/publication/pub.1044580340
244 rdf:type schema:CreativeWork
245 https://doi.org/10.1101/gad.12.8.1134 schema:sameAs https://app.dimensions.ai/details/publication/pub.1007603368
246 rdf:type schema:CreativeWork
247 https://doi.org/10.1101/gad.1780709 schema:sameAs https://app.dimensions.ai/details/publication/pub.1033256124
248 rdf:type schema:CreativeWork
249 https://doi.org/10.1101/gad.503108 schema:sameAs https://app.dimensions.ai/details/publication/pub.1017396089
250 rdf:type schema:CreativeWork
251 https://doi.org/10.1126/science.7855597 schema:sameAs https://app.dimensions.ai/details/publication/pub.1062649921
252 rdf:type schema:CreativeWork
253 https://doi.org/10.1128/mcb.14.12.8391 schema:sameAs https://app.dimensions.ai/details/publication/pub.1024726124
254 rdf:type schema:CreativeWork
255 https://doi.org/10.1128/mcb.24.21.9682-9694.2004 schema:sameAs https://app.dimensions.ai/details/publication/pub.1010422397
256 rdf:type schema:CreativeWork
257 https://doi.org/10.1128/mcb.25.11.4476-4487.2005 schema:sameAs https://app.dimensions.ai/details/publication/pub.1029059749
258 rdf:type schema:CreativeWork
259 https://doi.org/10.1128/mcb.25.13.5738-5751.2005 schema:sameAs https://app.dimensions.ai/details/publication/pub.1004857820
260 rdf:type schema:CreativeWork
261 https://doi.org/10.1128/mmbr.00020-08 schema:sameAs https://app.dimensions.ai/details/publication/pub.1041476570
262 rdf:type schema:CreativeWork
263 https://doi.org/10.1146/annurev.genet.38.072902.091500 schema:sameAs https://app.dimensions.ai/details/publication/pub.1028767256
264 rdf:type schema:CreativeWork
265 https://doi.org/10.1371/journal.pone.0004267 schema:sameAs https://app.dimensions.ai/details/publication/pub.1041896408
266 rdf:type schema:CreativeWork
267 https://www.grid.ac/institutes/grid.20861.3d schema:alternateName California Institute of Technology
268 schema:name Division of Biology, California Institute of Technology, Pasadena, California 91125, USA
269 rdf:type schema:Organization
270 https://www.grid.ac/institutes/grid.27860.3b schema:alternateName University of California, Davis
271 schema:name Department of Microbiology, University of California, Davis, Davis, California 95616-8665, USA
272 Department of Molecular and Cellular Biology, University of California, Davis, Davis, California 95616-8665, USA
273 rdf:type schema:Organization
 




Preview window. Press ESC to close (or click here)


...