ATP Hydrolysis by Mammalian RAD51 Has a Key Role during Homology-directed DNA Repair

Disruption of the gene encoding RAD51, the protein that catalyzes strand exchange during homologous recombination, leads to the accumulation of chromosome breaks and lethality in vertebrate cells. As RAD51 is implicated in BRCA1- and BRCA2-mediated tumor suppression as well as cellular viability, we...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:The Journal of biological chemistry 2002-06, Vol.277 (23), p.20185-20194
Hauptverfasser: Stark, Jeremy M., Hu, Peng, Pierce, Andrew J., Moynahan, Mary Ellen, Ellis, Nathan, Jasin, Maria
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 20194
container_issue 23
container_start_page 20185
container_title The Journal of biological chemistry
container_volume 277
creator Stark, Jeremy M.
Hu, Peng
Pierce, Andrew J.
Moynahan, Mary Ellen
Ellis, Nathan
Jasin, Maria
description Disruption of the gene encoding RAD51, the protein that catalyzes strand exchange during homologous recombination, leads to the accumulation of chromosome breaks and lethality in vertebrate cells. As RAD51 is implicated in BRCA1- and BRCA2-mediated tumor suppression as well as cellular viability, we have begun a functional analysis of a defined RAD51 mutation in mammalian cells. By using a dominant negative approach, we generated a mouse embryonic stem cell line that expresses an ATP hydrolysis-defective RAD51 protein, hRAD51-K133R, at comparable levels to the endogenous wild-type RAD51 protein, whose expression is retained in these cells. We found that these cells have increased sensitivity to the DNA-damaging agents mitomycin C and ionizing radiation and also exhibit a decreased rate of spontaneous sister-chromatid exchange. By using a reporter for the repair of a single chromosomal double-strand break, we also found that expression of the hRAD51-K133R protein specifically inhibits homology-directed double-strand break repair. Furthermore, expression of a BRC repeat from BRCA2, a peptide inhibitor of an early step necessary for strand exchange, exacerbates the inhibition of homology-directed repair in the hRAD51-K133R expressing cell line. Thus, ATP hydrolysis by RAD51 has a key role in various types of DNA repair in mammalian cells.
doi_str_mv 10.1074/jbc.M112132200
format Article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_71769975</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S002192582084856X</els_id><sourcerecordid>71769975</sourcerecordid><originalsourceid>FETCH-LOGICAL-c506t-9a041c303b27ffbcf9a95690f2153ad36e0c58ce4da85dea67fa22b98a29be03</originalsourceid><addsrcrecordid>eNqFkMFP2zAUxq0JNDrGlSPyAXFL8bPjJD5WwOg02KaqB26WY7-0Rkld7JYp_z1BqcQJ7V3e5fd9-vQj5BzYFFiZXz_XdvoIwEFwztgXMgFWiUxIeDoiE8Y4ZIrL6oR8S-mZDZcr-EpOABQXXPEJWc6Wf-m8dzG0ffKJ1j19NF1nWm82dDG7lUDnJlFDf2FPF6FF6vbRb1Z0HrrQhlWfOR_R7tDR298zusCt8fE7OW5Mm_Ds8E_J8sfd8maePfy5_3kze8isZMUuU4blYAUTNS-bpraNMkoWijUcpDBOFMisrCzmzlTSoSnKxnBeq8pwVSMTp-RqrN3G8LLHtNOdTxbb1mww7JMuoSyUKuV_QaiEkjkUAzgdQRtDShEbvY2-M7HXwPS7bz341h--h8DFoXlfd-g-8IPgAbgcgbVfrf8NqnTtg11jp3lZai40Z1C9L6xGDAddrx6jTtbjxuJoV7vgP5vwBpm4mBE</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>18395416</pqid></control><display><type>article</type><title>ATP Hydrolysis by Mammalian RAD51 Has a Key Role during Homology-directed DNA Repair</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Alma/SFX Local Collection</source><creator>Stark, Jeremy M. ; Hu, Peng ; Pierce, Andrew J. ; Moynahan, Mary Ellen ; Ellis, Nathan ; Jasin, Maria</creator><creatorcontrib>Stark, Jeremy M. ; Hu, Peng ; Pierce, Andrew J. ; Moynahan, Mary Ellen ; Ellis, Nathan ; Jasin, Maria</creatorcontrib><description>Disruption of the gene encoding RAD51, the protein that catalyzes strand exchange during homologous recombination, leads to the accumulation of chromosome breaks and lethality in vertebrate cells. As RAD51 is implicated in BRCA1- and BRCA2-mediated tumor suppression as well as cellular viability, we have begun a functional analysis of a defined RAD51 mutation in mammalian cells. By using a dominant negative approach, we generated a mouse embryonic stem cell line that expresses an ATP hydrolysis-defective RAD51 protein, hRAD51-K133R, at comparable levels to the endogenous wild-type RAD51 protein, whose expression is retained in these cells. We found that these cells have increased sensitivity to the DNA-damaging agents mitomycin C and ionizing radiation and also exhibit a decreased rate of spontaneous sister-chromatid exchange. By using a reporter for the repair of a single chromosomal double-strand break, we also found that expression of the hRAD51-K133R protein specifically inhibits homology-directed double-strand break repair. Furthermore, expression of a BRC repeat from BRCA2, a peptide inhibitor of an early step necessary for strand exchange, exacerbates the inhibition of homology-directed repair in the hRAD51-K133R expressing cell line. Thus, ATP hydrolysis by RAD51 has a key role in various types of DNA repair in mammalian cells.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M112132200</identifier><identifier>PMID: 11923292</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Adenosine Triphosphate - metabolism ; Animals ; Base Sequence ; DNA Damage ; DNA Primers ; DNA Repair ; DNA-Binding Proteins - metabolism ; Hydrolysis ; Mice ; Rad51 Recombinase ; Recombination, Genetic ; Sister Chromatid Exchange</subject><ispartof>The Journal of biological chemistry, 2002-06, Vol.277 (23), p.20185-20194</ispartof><rights>2002 © 2002 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c506t-9a041c303b27ffbcf9a95690f2153ad36e0c58ce4da85dea67fa22b98a29be03</citedby><cites>FETCH-LOGICAL-c506t-9a041c303b27ffbcf9a95690f2153ad36e0c58ce4da85dea67fa22b98a29be03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11923292$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Stark, Jeremy M.</creatorcontrib><creatorcontrib>Hu, Peng</creatorcontrib><creatorcontrib>Pierce, Andrew J.</creatorcontrib><creatorcontrib>Moynahan, Mary Ellen</creatorcontrib><creatorcontrib>Ellis, Nathan</creatorcontrib><creatorcontrib>Jasin, Maria</creatorcontrib><title>ATP Hydrolysis by Mammalian RAD51 Has a Key Role during Homology-directed DNA Repair</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Disruption of the gene encoding RAD51, the protein that catalyzes strand exchange during homologous recombination, leads to the accumulation of chromosome breaks and lethality in vertebrate cells. As RAD51 is implicated in BRCA1- and BRCA2-mediated tumor suppression as well as cellular viability, we have begun a functional analysis of a defined RAD51 mutation in mammalian cells. By using a dominant negative approach, we generated a mouse embryonic stem cell line that expresses an ATP hydrolysis-defective RAD51 protein, hRAD51-K133R, at comparable levels to the endogenous wild-type RAD51 protein, whose expression is retained in these cells. We found that these cells have increased sensitivity to the DNA-damaging agents mitomycin C and ionizing radiation and also exhibit a decreased rate of spontaneous sister-chromatid exchange. By using a reporter for the repair of a single chromosomal double-strand break, we also found that expression of the hRAD51-K133R protein specifically inhibits homology-directed double-strand break repair. Furthermore, expression of a BRC repeat from BRCA2, a peptide inhibitor of an early step necessary for strand exchange, exacerbates the inhibition of homology-directed repair in the hRAD51-K133R expressing cell line. Thus, ATP hydrolysis by RAD51 has a key role in various types of DNA repair in mammalian cells.</description><subject>Adenosine Triphosphate - metabolism</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>DNA Damage</subject><subject>DNA Primers</subject><subject>DNA Repair</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Hydrolysis</subject><subject>Mice</subject><subject>Rad51 Recombinase</subject><subject>Recombination, Genetic</subject><subject>Sister Chromatid Exchange</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMFP2zAUxq0JNDrGlSPyAXFL8bPjJD5WwOg02KaqB26WY7-0Rkld7JYp_z1BqcQJ7V3e5fd9-vQj5BzYFFiZXz_XdvoIwEFwztgXMgFWiUxIeDoiE8Y4ZIrL6oR8S-mZDZcr-EpOABQXXPEJWc6Wf-m8dzG0ffKJ1j19NF1nWm82dDG7lUDnJlFDf2FPF6FF6vbRb1Z0HrrQhlWfOR_R7tDR298zusCt8fE7OW5Mm_Ds8E_J8sfd8maePfy5_3kze8isZMUuU4blYAUTNS-bpraNMkoWijUcpDBOFMisrCzmzlTSoSnKxnBeq8pwVSMTp-RqrN3G8LLHtNOdTxbb1mww7JMuoSyUKuV_QaiEkjkUAzgdQRtDShEbvY2-M7HXwPS7bz341h--h8DFoXlfd-g-8IPgAbgcgbVfrf8NqnTtg11jp3lZai40Z1C9L6xGDAddrx6jTtbjxuJoV7vgP5vwBpm4mBE</recordid><startdate>20020607</startdate><enddate>20020607</enddate><creator>Stark, Jeremy M.</creator><creator>Hu, Peng</creator><creator>Pierce, Andrew J.</creator><creator>Moynahan, Mary Ellen</creator><creator>Ellis, Nathan</creator><creator>Jasin, Maria</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>20020607</creationdate><title>ATP Hydrolysis by Mammalian RAD51 Has a Key Role during Homology-directed DNA Repair</title><author>Stark, Jeremy M. ; Hu, Peng ; Pierce, Andrew J. ; Moynahan, Mary Ellen ; Ellis, Nathan ; Jasin, Maria</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c506t-9a041c303b27ffbcf9a95690f2153ad36e0c58ce4da85dea67fa22b98a29be03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Adenosine Triphosphate - metabolism</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>DNA Damage</topic><topic>DNA Primers</topic><topic>DNA Repair</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Hydrolysis</topic><topic>Mice</topic><topic>Rad51 Recombinase</topic><topic>Recombination, Genetic</topic><topic>Sister Chromatid Exchange</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Stark, Jeremy M.</creatorcontrib><creatorcontrib>Hu, Peng</creatorcontrib><creatorcontrib>Pierce, Andrew J.</creatorcontrib><creatorcontrib>Moynahan, Mary Ellen</creatorcontrib><creatorcontrib>Ellis, Nathan</creatorcontrib><creatorcontrib>Jasin, Maria</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Stark, Jeremy M.</au><au>Hu, Peng</au><au>Pierce, Andrew J.</au><au>Moynahan, Mary Ellen</au><au>Ellis, Nathan</au><au>Jasin, Maria</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>ATP Hydrolysis by Mammalian RAD51 Has a Key Role during Homology-directed DNA Repair</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2002-06-07</date><risdate>2002</risdate><volume>277</volume><issue>23</issue><spage>20185</spage><epage>20194</epage><pages>20185-20194</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Disruption of the gene encoding RAD51, the protein that catalyzes strand exchange during homologous recombination, leads to the accumulation of chromosome breaks and lethality in vertebrate cells. As RAD51 is implicated in BRCA1- and BRCA2-mediated tumor suppression as well as cellular viability, we have begun a functional analysis of a defined RAD51 mutation in mammalian cells. By using a dominant negative approach, we generated a mouse embryonic stem cell line that expresses an ATP hydrolysis-defective RAD51 protein, hRAD51-K133R, at comparable levels to the endogenous wild-type RAD51 protein, whose expression is retained in these cells. We found that these cells have increased sensitivity to the DNA-damaging agents mitomycin C and ionizing radiation and also exhibit a decreased rate of spontaneous sister-chromatid exchange. By using a reporter for the repair of a single chromosomal double-strand break, we also found that expression of the hRAD51-K133R protein specifically inhibits homology-directed double-strand break repair. Furthermore, expression of a BRC repeat from BRCA2, a peptide inhibitor of an early step necessary for strand exchange, exacerbates the inhibition of homology-directed repair in the hRAD51-K133R expressing cell line. Thus, ATP hydrolysis by RAD51 has a key role in various types of DNA repair in mammalian cells.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>11923292</pmid><doi>10.1074/jbc.M112132200</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0021-9258
ispartof The Journal of biological chemistry, 2002-06, Vol.277 (23), p.20185-20194
issn 0021-9258
1083-351X
language eng
recordid cdi_proquest_miscellaneous_71769975
source MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection
subjects Adenosine Triphosphate - metabolism
Animals
Base Sequence
DNA Damage
DNA Primers
DNA Repair
DNA-Binding Proteins - metabolism
Hydrolysis
Mice
Rad51 Recombinase
Recombination, Genetic
Sister Chromatid Exchange
title ATP Hydrolysis by Mammalian RAD51 Has a Key Role during Homology-directed DNA Repair
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T21%3A30%3A17IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=ATP%20Hydrolysis%20by%20Mammalian%20RAD51%20Has%20a%20Key%20Role%20during%20Homology-directed%20DNA%20Repair&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Stark,%20Jeremy%20M.&rft.date=2002-06-07&rft.volume=277&rft.issue=23&rft.spage=20185&rft.epage=20194&rft.pages=20185-20194&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.M112132200&rft_dat=%3Cproquest_cross%3E71769975%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=18395416&rft_id=info:pmid/11923292&rft_els_id=S002192582084856X&rfr_iscdi=true