Helicase-defective RuvB(D113E) promotes RuvAB-mediated branch migration in vitro
In Escherichia coli, the RuvA and RuvB proteins interact at Holliday junctions to promote branch migration leading to the formation of heteroduplex DNA. RuvA provides junction-binding specificity and RuvB drives ATP-dependent branch migration. Since RuvB contains sequence motifs characteristic of a...
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| Veröffentlicht in: | Journal of molecular biology 1999-10, Vol.293 (3), p.505-519 |
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| container_title | Journal of molecular biology |
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| creator | George, H Mézard, C Stasiak, A West, S C |
| description | In Escherichia coli, the RuvA and RuvB proteins interact at Holliday junctions to promote branch migration leading to the formation of heteroduplex DNA. RuvA provides junction-binding specificity and RuvB drives ATP-dependent branch migration. Since RuvB contains sequence motifs characteristic of a DNA helicase and RuvAB exhibit helicase activity in vitro, we have analysed the role of DNA unwinding in relation to branch migration. A mutant RuvB protein, RuvB(D113E), mutated in helicase motif II (the DExx box), has been purified to homogeneity. The mutant protein forms hexameric rings on DNA similar to those formed by wild-type protein and promotes branch migration in the presence of RuvA. However, RuvB(D113E) exhibits reduced ATPase activity and is severely compromised in its DNA helicase activity. Models for RuvAB-mediated branch migration that invoke only limited DNA unwinding activity are proposed. |
| doi_str_mv | 10.1006/jmbi.1999.3187 |
| format | Article |
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RuvA provides junction-binding specificity and RuvB drives ATP-dependent branch migration. Since RuvB contains sequence motifs characteristic of a DNA helicase and RuvAB exhibit helicase activity in vitro, we have analysed the role of DNA unwinding in relation to branch migration. A mutant RuvB protein, RuvB(D113E), mutated in helicase motif II (the DExx box), has been purified to homogeneity. The mutant protein forms hexameric rings on DNA similar to those formed by wild-type protein and promotes branch migration in the presence of RuvA. However, RuvB(D113E) exhibits reduced ATPase activity and is severely compromised in its DNA helicase activity. Models for RuvAB-mediated branch migration that invoke only limited DNA unwinding activity are proposed.</description><identifier>ISSN: 0022-2836</identifier><identifier>DOI: 10.1006/jmbi.1999.3187</identifier><identifier>PMID: 10543946</identifier><language>eng</language><publisher>England</publisher><subject>Adenosine Triphosphatases - chemistry ; Adenosine Triphosphatases - genetics ; Adenosine Triphosphatases - isolation & purification ; Adenosine Triphosphatases - metabolism ; Adenosine Triphosphate - metabolism ; Amino Acid Motifs ; Amino Acid Sequence ; Amino Acid Substitution ; Bacterial Proteins - chemistry ; Bacterial Proteins - genetics ; Bacterial Proteins - isolation & purification ; Bacterial Proteins - metabolism ; DNA - chemistry ; DNA - genetics ; DNA - metabolism ; DNA - ultrastructure ; DNA Helicases - chemistry ; DNA Helicases - genetics ; DNA Helicases - isolation & purification ; DNA Helicases - metabolism ; DNA, Single-Stranded - chemistry ; DNA, Single-Stranded - genetics ; DNA, Single-Stranded - metabolism ; DNA, Superhelical - chemistry ; DNA, Superhelical - genetics ; DNA, Superhelical - metabolism ; DNA-Binding Proteins - metabolism ; Escherichia coli ; Escherichia coli - enzymology ; Escherichia coli - genetics ; Escherichia coli Proteins ; Genes, Bacterial - genetics ; Genes, Bacterial - physiology ; Kinetics ; Models, Genetic ; Molecular Sequence Data ; Mutation ; Nucleic Acid Conformation ; Nucleic Acid Heteroduplexes - chemistry ; Nucleic Acid Heteroduplexes - genetics ; Nucleic Acid Heteroduplexes - metabolism ; Phenotype ; Recombination, Genetic - genetics ; Ultraviolet Rays</subject><ispartof>Journal of molecular biology, 1999-10, Vol.293 (3), p.505-519</ispartof><rights>Copyright 1999 Academic Press.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10543946$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>George, H</creatorcontrib><creatorcontrib>Mézard, C</creatorcontrib><creatorcontrib>Stasiak, A</creatorcontrib><creatorcontrib>West, S C</creatorcontrib><title>Helicase-defective RuvB(D113E) promotes RuvAB-mediated branch migration in vitro</title><title>Journal of molecular biology</title><addtitle>J Mol Biol</addtitle><description>In Escherichia coli, the RuvA and RuvB proteins interact at Holliday junctions to promote branch migration leading to the formation of heteroduplex DNA. RuvA provides junction-binding specificity and RuvB drives ATP-dependent branch migration. Since RuvB contains sequence motifs characteristic of a DNA helicase and RuvAB exhibit helicase activity in vitro, we have analysed the role of DNA unwinding in relation to branch migration. A mutant RuvB protein, RuvB(D113E), mutated in helicase motif II (the DExx box), has been purified to homogeneity. The mutant protein forms hexameric rings on DNA similar to those formed by wild-type protein and promotes branch migration in the presence of RuvA. However, RuvB(D113E) exhibits reduced ATPase activity and is severely compromised in its DNA helicase activity. Models for RuvAB-mediated branch migration that invoke only limited DNA unwinding activity are proposed.</description><subject>Adenosine Triphosphatases - chemistry</subject><subject>Adenosine Triphosphatases - genetics</subject><subject>Adenosine Triphosphatases - isolation & purification</subject><subject>Adenosine Triphosphatases - metabolism</subject><subject>Adenosine Triphosphate - metabolism</subject><subject>Amino Acid Motifs</subject><subject>Amino Acid Sequence</subject><subject>Amino Acid Substitution</subject><subject>Bacterial Proteins - chemistry</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - isolation & purification</subject><subject>Bacterial Proteins - metabolism</subject><subject>DNA - chemistry</subject><subject>DNA - genetics</subject><subject>DNA - metabolism</subject><subject>DNA - ultrastructure</subject><subject>DNA Helicases - chemistry</subject><subject>DNA Helicases - genetics</subject><subject>DNA Helicases - isolation & purification</subject><subject>DNA Helicases - metabolism</subject><subject>DNA, Single-Stranded - chemistry</subject><subject>DNA, Single-Stranded - genetics</subject><subject>DNA, Single-Stranded - metabolism</subject><subject>DNA, Superhelical - chemistry</subject><subject>DNA, Superhelical - genetics</subject><subject>DNA, Superhelical - metabolism</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Escherichia coli</subject><subject>Escherichia coli - enzymology</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli Proteins</subject><subject>Genes, Bacterial - genetics</subject><subject>Genes, Bacterial - physiology</subject><subject>Kinetics</subject><subject>Models, Genetic</subject><subject>Molecular Sequence Data</subject><subject>Mutation</subject><subject>Nucleic Acid Conformation</subject><subject>Nucleic Acid Heteroduplexes - chemistry</subject><subject>Nucleic Acid Heteroduplexes - genetics</subject><subject>Nucleic Acid Heteroduplexes - metabolism</subject><subject>Phenotype</subject><subject>Recombination, Genetic - genetics</subject><subject>Ultraviolet Rays</subject><issn>0022-2836</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkDtPwzAUhT2AaCmsjCgTgiHl-hE_xrYUilQJhGCO7NgBV3mU2InEvyeIMjNd6ejT0fkuQhcY5hiA3-5q4-dYKTWnWIojNAUgJCWS8gk6DWEHABll8gRNMGSMKsan6HnjKl_o4FLrSldEP7jkpR-W13cY0_VNsu_auo0u_ISLZVo763V0NjGdboqPpPbvnY6-bRLfJIOPXXuGjktdBXd-uDP0dr9-XW3S7dPD42qxTfeEypjyEjgwZS0DoixnAjgtqQIDJsuwwMoAjBZYAtcCRGmd4JKyQmUMKOeGztDVb--48LN3Iea1D4WrKt24tg85V4RkYpT4D8SCMiqZGMHLA9ib0TPfd77W3Vf-9yz6Dc5mZuo</recordid><startdate>19991029</startdate><enddate>19991029</enddate><creator>George, H</creator><creator>Mézard, C</creator><creator>Stasiak, A</creator><creator>West, S C</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7QL</scope><scope>7TM</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>19991029</creationdate><title>Helicase-defective RuvB(D113E) promotes RuvAB-mediated branch migration in vitro</title><author>George, H ; Mézard, C ; Stasiak, A ; West, S C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p238t-6f06049dd4029d647063f390b0b551719b003181806a707fde76834c9540366b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>Adenosine Triphosphatases - chemistry</topic><topic>Adenosine Triphosphatases - genetics</topic><topic>Adenosine Triphosphatases - isolation & purification</topic><topic>Adenosine Triphosphatases - metabolism</topic><topic>Adenosine Triphosphate - metabolism</topic><topic>Amino Acid Motifs</topic><topic>Amino Acid Sequence</topic><topic>Amino Acid Substitution</topic><topic>Bacterial Proteins - chemistry</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - isolation & purification</topic><topic>Bacterial Proteins - metabolism</topic><topic>DNA - chemistry</topic><topic>DNA - genetics</topic><topic>DNA - metabolism</topic><topic>DNA - ultrastructure</topic><topic>DNA Helicases - chemistry</topic><topic>DNA Helicases - genetics</topic><topic>DNA Helicases - isolation & purification</topic><topic>DNA Helicases - metabolism</topic><topic>DNA, Single-Stranded - chemistry</topic><topic>DNA, Single-Stranded - genetics</topic><topic>DNA, Single-Stranded - metabolism</topic><topic>DNA, Superhelical - chemistry</topic><topic>DNA, Superhelical - genetics</topic><topic>DNA, Superhelical - metabolism</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Escherichia coli</topic><topic>Escherichia coli - enzymology</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli Proteins</topic><topic>Genes, Bacterial - genetics</topic><topic>Genes, Bacterial - physiology</topic><topic>Kinetics</topic><topic>Models, Genetic</topic><topic>Molecular Sequence Data</topic><topic>Mutation</topic><topic>Nucleic Acid Conformation</topic><topic>Nucleic Acid Heteroduplexes - chemistry</topic><topic>Nucleic Acid Heteroduplexes - genetics</topic><topic>Nucleic Acid Heteroduplexes - metabolism</topic><topic>Phenotype</topic><topic>Recombination, Genetic - genetics</topic><topic>Ultraviolet Rays</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>George, H</creatorcontrib><creatorcontrib>Mézard, C</creatorcontrib><creatorcontrib>Stasiak, A</creatorcontrib><creatorcontrib>West, S C</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>George, H</au><au>Mézard, C</au><au>Stasiak, A</au><au>West, S C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Helicase-defective RuvB(D113E) promotes RuvAB-mediated branch migration in vitro</atitle><jtitle>Journal of molecular biology</jtitle><addtitle>J Mol Biol</addtitle><date>1999-10-29</date><risdate>1999</risdate><volume>293</volume><issue>3</issue><spage>505</spage><epage>519</epage><pages>505-519</pages><issn>0022-2836</issn><abstract>In Escherichia coli, the RuvA and RuvB proteins interact at Holliday junctions to promote branch migration leading to the formation of heteroduplex DNA. RuvA provides junction-binding specificity and RuvB drives ATP-dependent branch migration. Since RuvB contains sequence motifs characteristic of a DNA helicase and RuvAB exhibit helicase activity in vitro, we have analysed the role of DNA unwinding in relation to branch migration. A mutant RuvB protein, RuvB(D113E), mutated in helicase motif II (the DExx box), has been purified to homogeneity. The mutant protein forms hexameric rings on DNA similar to those formed by wild-type protein and promotes branch migration in the presence of RuvA. However, RuvB(D113E) exhibits reduced ATPase activity and is severely compromised in its DNA helicase activity. Models for RuvAB-mediated branch migration that invoke only limited DNA unwinding activity are proposed.</abstract><cop>England</cop><pmid>10543946</pmid><doi>10.1006/jmbi.1999.3187</doi><tpages>15</tpages></addata></record> |
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| subjects | Adenosine Triphosphatases - chemistry Adenosine Triphosphatases - genetics Adenosine Triphosphatases - isolation & purification Adenosine Triphosphatases - metabolism Adenosine Triphosphate - metabolism Amino Acid Motifs Amino Acid Sequence Amino Acid Substitution Bacterial Proteins - chemistry Bacterial Proteins - genetics Bacterial Proteins - isolation & purification Bacterial Proteins - metabolism DNA - chemistry DNA - genetics DNA - metabolism DNA - ultrastructure DNA Helicases - chemistry DNA Helicases - genetics DNA Helicases - isolation & purification DNA Helicases - metabolism DNA, Single-Stranded - chemistry DNA, Single-Stranded - genetics DNA, Single-Stranded - metabolism DNA, Superhelical - chemistry DNA, Superhelical - genetics DNA, Superhelical - metabolism DNA-Binding Proteins - metabolism Escherichia coli Escherichia coli - enzymology Escherichia coli - genetics Escherichia coli Proteins Genes, Bacterial - genetics Genes, Bacterial - physiology Kinetics Models, Genetic Molecular Sequence Data Mutation Nucleic Acid Conformation Nucleic Acid Heteroduplexes - chemistry Nucleic Acid Heteroduplexes - genetics Nucleic Acid Heteroduplexes - metabolism Phenotype Recombination, Genetic - genetics Ultraviolet Rays |
| title | Helicase-defective RuvB(D113E) promotes RuvAB-mediated branch migration in vitro |
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