The dynamics of Ku70/80 and DNA-PKcs at DSBs induced by ionizing radiation is dependent on the complexity of damage
DNA double-strand breaks (DSBs) are biologically one of the most important cellular lesions and possess varying degrees of chemical complexity. The notion that the repairability of more chemically complex DSBs is inefficient led to the concept that the extent of DSB complexity underlies the severity...
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| Veröffentlicht in: | Nucleic acids research 2012-11, Vol.40 (21), p.10821-10831 |
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| creator | Reynolds, Pamela Anderson, Jennifer A Harper, Jane V Hill, Mark A Botchway, Stanley W Parker, Anthony W O'Neill, Peter |
| description | DNA double-strand breaks (DSBs) are biologically one of the most important cellular lesions and possess varying degrees of chemical complexity. The notion that the repairability of more chemically complex DSBs is inefficient led to the concept that the extent of DSB complexity underlies the severity of the biological consequences. The repair of DSBs by non-homologous end joining (NHEJ) has been extensively studied but it remains unknown whether more complex DSBs require a different sub-set of NHEJ protein for their repair compared with simple DSBs. To address this, we have induced DSBs in fluorescently tagged mammalian cells (Ku80-EGFP, DNA-PKcs-YFP or XRCC4-GFP, key proteins in NHEJ) using ultra-soft X-rays (USX) or multi-photon near infrared (NIR) laser irradiation. We have shown in real-time that simple DSBs, induced by USX or NIR microbeam irradiation, are repaired rapidly involving Ku70/80 and XRCC4/Ligase IV/XLF. In contrast, DSBs with greater chemical complexity are repaired slowly involving not only Ku70/80 and XRCC4/Ligase IV/XLF but also DNA-PKcs. Ataxia telangiectasia-mutated inhibition only retards repair of the more chemically complex DSBs which require DNA-PKcs. In summary, the repair of DSBs by NHEJ is highly regulated with pathway choice and kinetics of repair dependent on the chemical complexity of the DSB. |
| doi_str_mv | 10.1093/nar/gks879 |
| format | Article |
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The notion that the repairability of more chemically complex DSBs is inefficient led to the concept that the extent of DSB complexity underlies the severity of the biological consequences. The repair of DSBs by non-homologous end joining (NHEJ) has been extensively studied but it remains unknown whether more complex DSBs require a different sub-set of NHEJ protein for their repair compared with simple DSBs. To address this, we have induced DSBs in fluorescently tagged mammalian cells (Ku80-EGFP, DNA-PKcs-YFP or XRCC4-GFP, key proteins in NHEJ) using ultra-soft X-rays (USX) or multi-photon near infrared (NIR) laser irradiation. We have shown in real-time that simple DSBs, induced by USX or NIR microbeam irradiation, are repaired rapidly involving Ku70/80 and XRCC4/Ligase IV/XLF. In contrast, DSBs with greater chemical complexity are repaired slowly involving not only Ku70/80 and XRCC4/Ligase IV/XLF but also DNA-PKcs. Ataxia telangiectasia-mutated inhibition only retards repair of the more chemically complex DSBs which require DNA-PKcs. In summary, the repair of DSBs by NHEJ is highly regulated with pathway choice and kinetics of repair dependent on the chemical complexity of the DSB.</description><identifier>ISSN: 0305-1048</identifier><identifier>EISSN: 1362-4962</identifier><identifier>DOI: 10.1093/nar/gks879</identifier><identifier>PMID: 23012265</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Animals ; Antigens, Nuclear - metabolism ; Ataxia ; Ataxia Telangiectasia Mutated Proteins ; BASIC BIOLOGICAL SCIENCES ; Cell Cycle Proteins - antagonists & inhibitors ; Cell Line ; DNA Breaks, Double-Stranded ; DNA damage ; DNA End-Joining Repair ; DNA-Activated Protein Kinase - metabolism ; DNA-Binding Proteins - antagonists & inhibitors ; DNA-Binding Proteins - metabolism ; DNA-dependent protein kinase ; Genome Integrity, Repair and ; I.R. radiation ; Ionizing radiation ; Kinetics ; Ku Autoantigen ; Lasers ; Mammalian cells ; Non-homologous end joining ; Protein-Serine-Threonine Kinases - antagonists & inhibitors ; Radiation, Ionizing ; Tumor Suppressor Proteins - antagonists & inhibitors</subject><ispartof>Nucleic acids research, 2012-11, Vol.40 (21), p.10821-10831</ispartof><rights>The Author(s) 2012. 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Ataxia telangiectasia-mutated inhibition only retards repair of the more chemically complex DSBs which require DNA-PKcs. In summary, the repair of DSBs by NHEJ is highly regulated with pathway choice and kinetics of repair dependent on the chemical complexity of the DSB.</description><subject>Animals</subject><subject>Antigens, Nuclear - metabolism</subject><subject>Ataxia</subject><subject>Ataxia Telangiectasia Mutated Proteins</subject><subject>BASIC BIOLOGICAL SCIENCES</subject><subject>Cell Cycle Proteins - antagonists & inhibitors</subject><subject>Cell Line</subject><subject>DNA Breaks, Double-Stranded</subject><subject>DNA damage</subject><subject>DNA End-Joining Repair</subject><subject>DNA-Activated Protein Kinase - metabolism</subject><subject>DNA-Binding Proteins - antagonists & inhibitors</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>DNA-dependent protein kinase</subject><subject>Genome Integrity, Repair and</subject><subject>I.R. radiation</subject><subject>Ionizing radiation</subject><subject>Kinetics</subject><subject>Ku Autoantigen</subject><subject>Lasers</subject><subject>Mammalian cells</subject><subject>Non-homologous end joining</subject><subject>Protein-Serine-Threonine Kinases - antagonists & inhibitors</subject><subject>Radiation, Ionizing</subject><subject>Tumor Suppressor Proteins - antagonists & inhibitors</subject><issn>0305-1048</issn><issn>1362-4962</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU1vEzEQhi0EoqFw4QcgixNCWjL-Wu9ekErLl1oBEuVsee1JYsjaYe1FhF-Po5QKbpys8Tx65515CXnM4AWDXiyjnZbrb7nT_R2yYKLljexbfpcsQIBqGMjuhDzI-SsAk0zJ--SEC2Cct2pB8vUGqd9HOwaXaVrRy1nDsgNqo6cXH86aT5f13xZ68flVpiH62aGnw56GFMOvENd0sj7YUksaMvW4w-gxFlrrUqVdGndb_BnK_iDu7WjX-JDcW9ltxkc37yn58ub19fm75urj2_fnZ1eNUyBLIzSC5avWo-6Rc9G1yH0HYuiFd0xrP0gL3ql-4KrtrfVcolaAKDS0YmjFKXl51N3Nw4jeVVuT3ZrdFEY77U2ywfzbiWFj1umHEaoerWdV4OlRIOUSTHahoNu4FCO6YlgPUskD9OxmypS-z5iLGUN2uN3aiGnOhnGlqy3V6f9AOdRVpIaKPj-ibko5T7i6tc3AHFI3NXVzTL3CT_5e9Bb9E7P4DSesqB4</recordid><startdate>20121101</startdate><enddate>20121101</enddate><creator>Reynolds, Pamela</creator><creator>Anderson, Jennifer A</creator><creator>Harper, Jane V</creator><creator>Hill, Mark A</creator><creator>Botchway, Stanley W</creator><creator>Parker, Anthony W</creator><creator>O'Neill, Peter</creator><general>Oxford University Press</general><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>7X8</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>OIOZB</scope><scope>OTOTI</scope><scope>5PM</scope></search><sort><creationdate>20121101</creationdate><title>The dynamics of Ku70/80 and DNA-PKcs at DSBs induced by ionizing radiation is dependent on the complexity of damage</title><author>Reynolds, Pamela ; Anderson, Jennifer A ; Harper, Jane V ; Hill, Mark A ; Botchway, Stanley W ; Parker, Anthony W ; O'Neill, Peter</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c504t-37e0a2f6de79e22386e2d803b93dc177db4a0dc59b2569aad24e750ee37063b63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Animals</topic><topic>Antigens, Nuclear - metabolism</topic><topic>Ataxia</topic><topic>Ataxia Telangiectasia Mutated Proteins</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>Cell Cycle Proteins - antagonists & inhibitors</topic><topic>Cell Line</topic><topic>DNA Breaks, Double-Stranded</topic><topic>DNA damage</topic><topic>DNA End-Joining Repair</topic><topic>DNA-Activated Protein Kinase - metabolism</topic><topic>DNA-Binding Proteins - antagonists & inhibitors</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>DNA-dependent protein kinase</topic><topic>Genome Integrity, Repair and</topic><topic>I.R. radiation</topic><topic>Ionizing radiation</topic><topic>Kinetics</topic><topic>Ku Autoantigen</topic><topic>Lasers</topic><topic>Mammalian cells</topic><topic>Non-homologous end joining</topic><topic>Protein-Serine-Threonine Kinases - antagonists & inhibitors</topic><topic>Radiation, Ionizing</topic><topic>Tumor Suppressor Proteins - antagonists & inhibitors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Reynolds, Pamela</creatorcontrib><creatorcontrib>Anderson, Jennifer A</creatorcontrib><creatorcontrib>Harper, Jane V</creatorcontrib><creatorcontrib>Hill, Mark A</creatorcontrib><creatorcontrib>Botchway, Stanley W</creatorcontrib><creatorcontrib>Parker, Anthony W</creatorcontrib><creatorcontrib>O'Neill, Peter</creatorcontrib><creatorcontrib>Univ. of Oxford (United Kingdom)</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>OSTI.GOV - Hybrid</collection><collection>OSTI.GOV</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nucleic acids research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Reynolds, Pamela</au><au>Anderson, Jennifer A</au><au>Harper, Jane V</au><au>Hill, Mark A</au><au>Botchway, Stanley W</au><au>Parker, Anthony W</au><au>O'Neill, Peter</au><aucorp>Univ. of Oxford (United Kingdom)</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The dynamics of Ku70/80 and DNA-PKcs at DSBs induced by ionizing radiation is dependent on the complexity of damage</atitle><jtitle>Nucleic acids research</jtitle><addtitle>Nucleic Acids Res</addtitle><date>2012-11-01</date><risdate>2012</risdate><volume>40</volume><issue>21</issue><spage>10821</spage><epage>10831</epage><pages>10821-10831</pages><issn>0305-1048</issn><eissn>1362-4962</eissn><abstract>DNA double-strand breaks (DSBs) are biologically one of the most important cellular lesions and possess varying degrees of chemical complexity. The notion that the repairability of more chemically complex DSBs is inefficient led to the concept that the extent of DSB complexity underlies the severity of the biological consequences. The repair of DSBs by non-homologous end joining (NHEJ) has been extensively studied but it remains unknown whether more complex DSBs require a different sub-set of NHEJ protein for their repair compared with simple DSBs. To address this, we have induced DSBs in fluorescently tagged mammalian cells (Ku80-EGFP, DNA-PKcs-YFP or XRCC4-GFP, key proteins in NHEJ) using ultra-soft X-rays (USX) or multi-photon near infrared (NIR) laser irradiation. We have shown in real-time that simple DSBs, induced by USX or NIR microbeam irradiation, are repaired rapidly involving Ku70/80 and XRCC4/Ligase IV/XLF. In contrast, DSBs with greater chemical complexity are repaired slowly involving not only Ku70/80 and XRCC4/Ligase IV/XLF but also DNA-PKcs. Ataxia telangiectasia-mutated inhibition only retards repair of the more chemically complex DSBs which require DNA-PKcs. In summary, the repair of DSBs by NHEJ is highly regulated with pathway choice and kinetics of repair dependent on the chemical complexity of the DSB.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>23012265</pmid><doi>10.1093/nar/gks879</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Antigens, Nuclear - metabolism Ataxia Ataxia Telangiectasia Mutated Proteins BASIC BIOLOGICAL SCIENCES Cell Cycle Proteins - antagonists & inhibitors Cell Line DNA Breaks, Double-Stranded DNA damage DNA End-Joining Repair DNA-Activated Protein Kinase - metabolism DNA-Binding Proteins - antagonists & inhibitors DNA-Binding Proteins - metabolism DNA-dependent protein kinase Genome Integrity, Repair and I.R. radiation Ionizing radiation Kinetics Ku Autoantigen Lasers Mammalian cells Non-homologous end joining Protein-Serine-Threonine Kinases - antagonists & inhibitors Radiation, Ionizing Tumor Suppressor Proteins - antagonists & inhibitors |
| title | The dynamics of Ku70/80 and DNA-PKcs at DSBs induced by ionizing radiation is dependent on the complexity of damage |
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