Polymerase theta-helicase promotes end joining by stripping single-stranded DNA-binding proteins and bridging DNA ends
Abstract Homologous recombination-deficient cancers rely on DNA polymerase Theta (Polθ)-Mediated End Joining (TMEJ), an alternative double-strand break repair pathway. Polθ is the only vertebrate polymerase that encodes an N-terminal superfamily 2 (SF2) helicase domain, but the role of this helicase...
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| Veröffentlicht in: | Nucleic acids research 2022-04, Vol.50 (7), p.3911-3921 |
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| creator | Schaub, Jeffrey M Soniat, Michael M Finkelstein, Ilya J |
| description | Abstract
Homologous recombination-deficient cancers rely on DNA polymerase Theta (Polθ)-Mediated End Joining (TMEJ), an alternative double-strand break repair pathway. Polθ is the only vertebrate polymerase that encodes an N-terminal superfamily 2 (SF2) helicase domain, but the role of this helicase domain in TMEJ remains unclear. Using single-molecule imaging, we demonstrate that Polθ-helicase (Polθ-h) is a highly processive single-stranded DNA (ssDNA) motor protein that can efficiently strip Replication Protein A (RPA) from ssDNA. Polθ-h also has a limited capacity for disassembling RAD51 filaments but is not processive on double-stranded DNA. Polθ-h can bridge two non-complementary DNA strands in trans. PARylation of Polθ-h by PARP-1 resolves these DNA bridges. We conclude that Polθ-h removes RPA and RAD51 filaments and mediates bridging of DNA overhangs to aid in polymerization by the Polθ polymerase domain. |
| doi_str_mv | 10.1093/nar/gkac119 |
| format | Article |
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Homologous recombination-deficient cancers rely on DNA polymerase Theta (Polθ)-Mediated End Joining (TMEJ), an alternative double-strand break repair pathway. Polθ is the only vertebrate polymerase that encodes an N-terminal superfamily 2 (SF2) helicase domain, but the role of this helicase domain in TMEJ remains unclear. Using single-molecule imaging, we demonstrate that Polθ-helicase (Polθ-h) is a highly processive single-stranded DNA (ssDNA) motor protein that can efficiently strip Replication Protein A (RPA) from ssDNA. Polθ-h also has a limited capacity for disassembling RAD51 filaments but is not processive on double-stranded DNA. Polθ-h can bridge two non-complementary DNA strands in trans. PARylation of Polθ-h by PARP-1 resolves these DNA bridges. We conclude that Polθ-h removes RPA and RAD51 filaments and mediates bridging of DNA overhangs to aid in polymerization by the Polθ polymerase domain.</description><identifier>ISSN: 0305-1048</identifier><identifier>EISSN: 1362-4962</identifier><identifier>DOI: 10.1093/nar/gkac119</identifier><identifier>PMID: 35357490</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>DNA - chemistry ; DNA Breaks, Double-Stranded ; DNA End-Joining Repair ; DNA Helicases - genetics ; DNA, Single-Stranded - genetics ; DNA-Binding Proteins - genetics ; DNA-Binding Proteins - metabolism ; Genome Integrity, Repair and ; Replication Protein A - genetics ; Replication Protein A - metabolism</subject><ispartof>Nucleic acids research, 2022-04, Vol.50 (7), p.3911-3921</ispartof><rights>The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research. 2022</rights><rights>The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c478t-d2e75fa1ef7f2bc1189918b4df59f85b5c1615bfd0b0ab5dedcf2e09c5a3b1fa3</citedby><cites>FETCH-LOGICAL-c478t-d2e75fa1ef7f2bc1189918b4df59f85b5c1615bfd0b0ab5dedcf2e09c5a3b1fa3</cites><orcidid>0000-0002-9371-2431</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9023281/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9023281/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,1598,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35357490$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Schaub, Jeffrey M</creatorcontrib><creatorcontrib>Soniat, Michael M</creatorcontrib><creatorcontrib>Finkelstein, Ilya J</creatorcontrib><title>Polymerase theta-helicase promotes end joining by stripping single-stranded DNA-binding proteins and bridging DNA ends</title><title>Nucleic acids research</title><addtitle>Nucleic Acids Res</addtitle><description>Abstract
Homologous recombination-deficient cancers rely on DNA polymerase Theta (Polθ)-Mediated End Joining (TMEJ), an alternative double-strand break repair pathway. Polθ is the only vertebrate polymerase that encodes an N-terminal superfamily 2 (SF2) helicase domain, but the role of this helicase domain in TMEJ remains unclear. Using single-molecule imaging, we demonstrate that Polθ-helicase (Polθ-h) is a highly processive single-stranded DNA (ssDNA) motor protein that can efficiently strip Replication Protein A (RPA) from ssDNA. Polθ-h also has a limited capacity for disassembling RAD51 filaments but is not processive on double-stranded DNA. Polθ-h can bridge two non-complementary DNA strands in trans. PARylation of Polθ-h by PARP-1 resolves these DNA bridges. We conclude that Polθ-h removes RPA and RAD51 filaments and mediates bridging of DNA overhangs to aid in polymerization by the Polθ polymerase domain.</description><subject>DNA - chemistry</subject><subject>DNA Breaks, Double-Stranded</subject><subject>DNA End-Joining Repair</subject><subject>DNA Helicases - genetics</subject><subject>DNA, Single-Stranded - genetics</subject><subject>DNA-Binding Proteins - genetics</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Genome Integrity, Repair and</subject><subject>Replication Protein A - genetics</subject><subject>Replication Protein A - metabolism</subject><issn>0305-1048</issn><issn>1362-4962</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>TOX</sourceid><sourceid>EIF</sourceid><recordid>eNp9kc9LwzAUx4MoOqcn79KTCFKXtM3aXIThbxjqQc8haV-2zDapSSvsvzdlU_TiJeG97yff98IXoROCLwlm6cQIN1m8i5IQtoNGJJ0mccamyS4a4RTTmOCsOECH3q8wJhmh2T46SGlK84zhEfp8sfW6ASc8RN0SOhEvodblULbONrYDH4GpopXVRptFJNeR75xu26Hw4aghDg1hKqiim6dZLLWpBi287kAbHwUpkk5Xi6EbiMHOH6E9JWoPx9t7jN7ubl-vH-L58_3j9Wwel1ledHGVQE6VIKBylcjww4IxUsisUpSpgkpakimhUlVYYiFp2KFUCWBWUpFKokQ6Rlcb37aXTVDBhF1r3jrdCLfmVmj-VzF6yRf2kzOcpElBgsH51sDZjx58xxvtS6hrYcD2nifTjBa0yCkL6MUGLZ313oH6GUMwH5LiISm-TSrQp783-2G_ownA2Qawffuv0xfdRaGP</recordid><startdate>20220422</startdate><enddate>20220422</enddate><creator>Schaub, Jeffrey M</creator><creator>Soniat, Michael M</creator><creator>Finkelstein, Ilya J</creator><general>Oxford University Press</general><scope>TOX</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>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-9371-2431</orcidid></search><sort><creationdate>20220422</creationdate><title>Polymerase theta-helicase promotes end joining by stripping single-stranded DNA-binding proteins and bridging DNA ends</title><author>Schaub, Jeffrey M ; Soniat, Michael M ; Finkelstein, Ilya J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c478t-d2e75fa1ef7f2bc1189918b4df59f85b5c1615bfd0b0ab5dedcf2e09c5a3b1fa3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>DNA - chemistry</topic><topic>DNA Breaks, Double-Stranded</topic><topic>DNA End-Joining Repair</topic><topic>DNA Helicases - genetics</topic><topic>DNA, Single-Stranded - genetics</topic><topic>DNA-Binding Proteins - genetics</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Genome Integrity, Repair and</topic><topic>Replication Protein A - genetics</topic><topic>Replication Protein A - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Schaub, Jeffrey M</creatorcontrib><creatorcontrib>Soniat, Michael M</creatorcontrib><creatorcontrib>Finkelstein, Ilya J</creatorcontrib><collection>Oxford Journals Open Access Collection</collection><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>PubMed Central (Full Participant titles)</collection><jtitle>Nucleic acids research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Schaub, Jeffrey M</au><au>Soniat, Michael M</au><au>Finkelstein, Ilya J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Polymerase theta-helicase promotes end joining by stripping single-stranded DNA-binding proteins and bridging DNA ends</atitle><jtitle>Nucleic acids research</jtitle><addtitle>Nucleic Acids Res</addtitle><date>2022-04-22</date><risdate>2022</risdate><volume>50</volume><issue>7</issue><spage>3911</spage><epage>3921</epage><pages>3911-3921</pages><issn>0305-1048</issn><eissn>1362-4962</eissn><abstract>Abstract
Homologous recombination-deficient cancers rely on DNA polymerase Theta (Polθ)-Mediated End Joining (TMEJ), an alternative double-strand break repair pathway. Polθ is the only vertebrate polymerase that encodes an N-terminal superfamily 2 (SF2) helicase domain, but the role of this helicase domain in TMEJ remains unclear. Using single-molecule imaging, we demonstrate that Polθ-helicase (Polθ-h) is a highly processive single-stranded DNA (ssDNA) motor protein that can efficiently strip Replication Protein A (RPA) from ssDNA. Polθ-h also has a limited capacity for disassembling RAD51 filaments but is not processive on double-stranded DNA. Polθ-h can bridge two non-complementary DNA strands in trans. PARylation of Polθ-h by PARP-1 resolves these DNA bridges. We conclude that Polθ-h removes RPA and RAD51 filaments and mediates bridging of DNA overhangs to aid in polymerization by the Polθ polymerase domain.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>35357490</pmid><doi>10.1093/nar/gkac119</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0002-9371-2431</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | DNA - chemistry DNA Breaks, Double-Stranded DNA End-Joining Repair DNA Helicases - genetics DNA, Single-Stranded - genetics DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism Genome Integrity, Repair and Replication Protein A - genetics Replication Protein A - metabolism |
| title | Polymerase theta-helicase promotes end joining by stripping single-stranded DNA-binding proteins and bridging DNA ends |
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