Mutational analyses of the thermostable NAD +-dependent DNA ligase from Thermus filiformis
The crystal structure of NAD +-dependent DNA ligase from Thermus filiformis ( Tfi) revealed that the protein comprised four structural domains. In order to investigate the biochemical activities of these domains, seven deletion mutants were constructed from the Tfi DNA ligase. The mutants Tfi-M1 (re...
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| Veröffentlicht in: | FEMS microbiology letters 2004-08, Vol.237 (1), p.111-118 |
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| creator | Jeon, Hyo Jeong Shin, Hea-Jin Choi, Jeong Jin Hoe, Hyang-Sook Kim, Hyun-Kyu Suh, Se Won Kwon, Suk-Tae |
| description | The crystal structure of NAD
+-dependent DNA ligase from
Thermus filiformis (
Tfi) revealed that the protein comprised four structural domains. In order to investigate the biochemical activities of these domains, seven deletion mutants were constructed from the
Tfi DNA ligase. The mutants
Tfi-M1 (residues 1–581),
Tfi-M2 (residues 1–448),
Tfi-M3 (residues 1–403) and
Tfi-M4 (residues 1–314) showed the same adenylation activity as that of wild-type. This result indicates that only the adenylation domain (domain 1) is essential for the formation of enzyme–AMP complex. It was found that the zinc finger and helix-hairpin-helix (HhH) motif domain (domain 3) and the oligomer binding (OB)-fold domain (domain 2) are important for the formation of enzyme–DNA complex. The mutant
Tfi-M1 alone showed the activities for in vitro nick-closing and in vivo complementation in
Escherichia coli as those of wild-type. These results indicate that the BRCT domain (domain 4) of
Tfi DNA ligase is not essential for the enzyme activity. The enzymatic properties of
Tfi-M1 mutant (deleted the BRCT domain) were slightly different from those of wild-type and the nick-closing activity of
Tfi-M1 mutant was approximately 50% compared with that of wild-type. |
| doi_str_mv | 10.1016/j.femsle.2004.06.018 |
| format | Article |
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+-dependent DNA ligase from
Thermus filiformis (
Tfi) revealed that the protein comprised four structural domains. In order to investigate the biochemical activities of these domains, seven deletion mutants were constructed from the
Tfi DNA ligase. The mutants
Tfi-M1 (residues 1–581),
Tfi-M2 (residues 1–448),
Tfi-M3 (residues 1–403) and
Tfi-M4 (residues 1–314) showed the same adenylation activity as that of wild-type. This result indicates that only the adenylation domain (domain 1) is essential for the formation of enzyme–AMP complex. It was found that the zinc finger and helix-hairpin-helix (HhH) motif domain (domain 3) and the oligomer binding (OB)-fold domain (domain 2) are important for the formation of enzyme–DNA complex. The mutant
Tfi-M1 alone showed the activities for in vitro nick-closing and in vivo complementation in
Escherichia coli as those of wild-type. These results indicate that the BRCT domain (domain 4) of
Tfi DNA ligase is not essential for the enzyme activity. The enzymatic properties of
Tfi-M1 mutant (deleted the BRCT domain) were slightly different from those of wild-type and the nick-closing activity of
Tfi-M1 mutant was approximately 50% compared with that of wild-type.</description><identifier>ISSN: 0378-1097</identifier><identifier>EISSN: 1574-6968</identifier><identifier>DOI: 10.1016/j.femsle.2004.06.018</identifier><identifier>PMID: 15268945</identifier><identifier>CODEN: FMLED7</identifier><language>eng</language><publisher>Oxford: Elsevier B.V</publisher><subject>Adenosine Monophosphate - metabolism ; Bacteriology ; Biological and medical sciences ; Cloning, Molecular ; Coenzymes - pharmacology ; DNA binding activity ; DNA Ligases - genetics ; DNA Ligases - isolation & purification ; DNA Ligases - metabolism ; DNA Mutational Analysis ; DNA-Binding Proteins - genetics ; DNA-Binding Proteins - physiology ; Enzyme Stability ; Escherichia coli - genetics ; Escherichia coli - growth & development ; Escherichia coli - metabolism ; Fundamental and applied biological sciences. Psychology ; Genetic Complementation Test ; Helix-Turn-Helix Motifs - genetics ; Helix-Turn-Helix Motifs - physiology ; Hydrogen-Ion Concentration ; Microbiology ; Miscellaneous ; Nick-closing activity ; Protein Structure, Tertiary ; Recombinant Proteins - genetics ; Recombinant Proteins - isolation & purification ; Recombinant Proteins - metabolism ; Self-adenylation ; Sequence Deletion ; Temperature ; Thermus - enzymology ; Thermus filiformis ( Tfi) DNA ligase ; Time Factors ; Zinc Fingers - genetics ; Zinc Fingers - physiology</subject><ispartof>FEMS microbiology letters, 2004-08, Vol.237 (1), p.111-118</ispartof><rights>2004 Federation of European Microbiological Societies</rights><rights>2005 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c343t-2a7425233a23d490586541cd8c01bbf22b19809ce46f195d3c2868d2f0ab1b333</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15954514$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15268945$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jeon, Hyo Jeong</creatorcontrib><creatorcontrib>Shin, Hea-Jin</creatorcontrib><creatorcontrib>Choi, Jeong Jin</creatorcontrib><creatorcontrib>Hoe, Hyang-Sook</creatorcontrib><creatorcontrib>Kim, Hyun-Kyu</creatorcontrib><creatorcontrib>Suh, Se Won</creatorcontrib><creatorcontrib>Kwon, Suk-Tae</creatorcontrib><title>Mutational analyses of the thermostable NAD +-dependent DNA ligase from Thermus filiformis</title><title>FEMS microbiology letters</title><addtitle>FEMS Microbiol Lett</addtitle><description>The crystal structure of NAD
+-dependent DNA ligase from
Thermus filiformis (
Tfi) revealed that the protein comprised four structural domains. In order to investigate the biochemical activities of these domains, seven deletion mutants were constructed from the
Tfi DNA ligase. The mutants
Tfi-M1 (residues 1–581),
Tfi-M2 (residues 1–448),
Tfi-M3 (residues 1–403) and
Tfi-M4 (residues 1–314) showed the same adenylation activity as that of wild-type. This result indicates that only the adenylation domain (domain 1) is essential for the formation of enzyme–AMP complex. It was found that the zinc finger and helix-hairpin-helix (HhH) motif domain (domain 3) and the oligomer binding (OB)-fold domain (domain 2) are important for the formation of enzyme–DNA complex. The mutant
Tfi-M1 alone showed the activities for in vitro nick-closing and in vivo complementation in
Escherichia coli as those of wild-type. These results indicate that the BRCT domain (domain 4) of
Tfi DNA ligase is not essential for the enzyme activity. The enzymatic properties of
Tfi-M1 mutant (deleted the BRCT domain) were slightly different from those of wild-type and the nick-closing activity of
Tfi-M1 mutant was approximately 50% compared with that of wild-type.</description><subject>Adenosine Monophosphate - metabolism</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Cloning, Molecular</subject><subject>Coenzymes - pharmacology</subject><subject>DNA binding activity</subject><subject>DNA Ligases - genetics</subject><subject>DNA Ligases - isolation & purification</subject><subject>DNA Ligases - metabolism</subject><subject>DNA Mutational Analysis</subject><subject>DNA-Binding Proteins - genetics</subject><subject>DNA-Binding Proteins - physiology</subject><subject>Enzyme Stability</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - growth & development</subject><subject>Escherichia coli - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetic Complementation Test</subject><subject>Helix-Turn-Helix Motifs - genetics</subject><subject>Helix-Turn-Helix Motifs - physiology</subject><subject>Hydrogen-Ion Concentration</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>Nick-closing activity</subject><subject>Protein Structure, Tertiary</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Recombinant Proteins - metabolism</subject><subject>Self-adenylation</subject><subject>Sequence Deletion</subject><subject>Temperature</subject><subject>Thermus - enzymology</subject><subject>Thermus filiformis ( Tfi) DNA ligase</subject><subject>Time Factors</subject><subject>Zinc Fingers - genetics</subject><subject>Zinc Fingers - physiology</subject><issn>0378-1097</issn><issn>1574-6968</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpN0U1v1DAQBmALgei28A8Q8gUuKKm_Y1-QVi2lSG25lAsXy7HH4JWTLHaC1H9PVrtIPXh8eTSjmRehd5S0lFB1uWsjDDVDywgRLVEtofoF2lDZiUYZpV-iDeGdbigx3Rk6r3VHVsiIeo3OqGRKGyE36Of9Mrs5TaPL2K3lqULFU8Tzbzi8Mkx1dn0G_LC9xp-aAHsYA4wzvn7Y4px-uQo4lmnAjwe8VBxTTnEqQ6pv0KvocoW3p_8C_bj58nh129x9__rtanvXeC743DDXCSYZ547xIAyRWklBfdCe0L6PjPXUaGI8CBWpkYF7ppUOLBLX055zfoE-Hvvuy_RngTrbdbiHnN0I01KtUh1XUpsVvj_BpR8g2H1JgytP9v81VvDhBFz1LsfiRp_qM2ekkFSs7vPRwbrW3wTFVp9g9BBSAT_bMCVLiT3EZHf2GJM9xGSJsmtM_B_yToSU</recordid><startdate>20040801</startdate><enddate>20040801</enddate><creator>Jeon, Hyo Jeong</creator><creator>Shin, Hea-Jin</creator><creator>Choi, Jeong Jin</creator><creator>Hoe, Hyang-Sook</creator><creator>Kim, Hyun-Kyu</creator><creator>Suh, Se Won</creator><creator>Kwon, Suk-Tae</creator><general>Elsevier B.V</general><general>Blackwell</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>20040801</creationdate><title>Mutational analyses of the thermostable NAD +-dependent DNA ligase from Thermus filiformis</title><author>Jeon, Hyo Jeong ; Shin, Hea-Jin ; Choi, Jeong Jin ; Hoe, Hyang-Sook ; Kim, Hyun-Kyu ; Suh, Se Won ; Kwon, Suk-Tae</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c343t-2a7425233a23d490586541cd8c01bbf22b19809ce46f195d3c2868d2f0ab1b333</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Adenosine Monophosphate - metabolism</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Cloning, Molecular</topic><topic>Coenzymes - pharmacology</topic><topic>DNA binding activity</topic><topic>DNA Ligases - genetics</topic><topic>DNA Ligases - isolation & purification</topic><topic>DNA Ligases - metabolism</topic><topic>DNA Mutational Analysis</topic><topic>DNA-Binding Proteins - genetics</topic><topic>DNA-Binding Proteins - physiology</topic><topic>Enzyme Stability</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - growth & development</topic><topic>Escherichia coli - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genetic Complementation Test</topic><topic>Helix-Turn-Helix Motifs - genetics</topic><topic>Helix-Turn-Helix Motifs - physiology</topic><topic>Hydrogen-Ion Concentration</topic><topic>Microbiology</topic><topic>Miscellaneous</topic><topic>Nick-closing activity</topic><topic>Protein Structure, Tertiary</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Recombinant Proteins - metabolism</topic><topic>Self-adenylation</topic><topic>Sequence Deletion</topic><topic>Temperature</topic><topic>Thermus - enzymology</topic><topic>Thermus filiformis ( Tfi) DNA ligase</topic><topic>Time Factors</topic><topic>Zinc Fingers - genetics</topic><topic>Zinc Fingers - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jeon, Hyo Jeong</creatorcontrib><creatorcontrib>Shin, Hea-Jin</creatorcontrib><creatorcontrib>Choi, Jeong Jin</creatorcontrib><creatorcontrib>Hoe, Hyang-Sook</creatorcontrib><creatorcontrib>Kim, Hyun-Kyu</creatorcontrib><creatorcontrib>Suh, Se Won</creatorcontrib><creatorcontrib>Kwon, Suk-Tae</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>FEMS microbiology letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jeon, Hyo Jeong</au><au>Shin, Hea-Jin</au><au>Choi, Jeong Jin</au><au>Hoe, Hyang-Sook</au><au>Kim, Hyun-Kyu</au><au>Suh, Se Won</au><au>Kwon, Suk-Tae</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Mutational analyses of the thermostable NAD +-dependent DNA ligase from Thermus filiformis</atitle><jtitle>FEMS microbiology letters</jtitle><addtitle>FEMS Microbiol Lett</addtitle><date>2004-08-01</date><risdate>2004</risdate><volume>237</volume><issue>1</issue><spage>111</spage><epage>118</epage><pages>111-118</pages><issn>0378-1097</issn><eissn>1574-6968</eissn><coden>FMLED7</coden><abstract>The crystal structure of NAD
+-dependent DNA ligase from
Thermus filiformis (
Tfi) revealed that the protein comprised four structural domains. In order to investigate the biochemical activities of these domains, seven deletion mutants were constructed from the
Tfi DNA ligase. The mutants
Tfi-M1 (residues 1–581),
Tfi-M2 (residues 1–448),
Tfi-M3 (residues 1–403) and
Tfi-M4 (residues 1–314) showed the same adenylation activity as that of wild-type. This result indicates that only the adenylation domain (domain 1) is essential for the formation of enzyme–AMP complex. It was found that the zinc finger and helix-hairpin-helix (HhH) motif domain (domain 3) and the oligomer binding (OB)-fold domain (domain 2) are important for the formation of enzyme–DNA complex. The mutant
Tfi-M1 alone showed the activities for in vitro nick-closing and in vivo complementation in
Escherichia coli as those of wild-type. These results indicate that the BRCT domain (domain 4) of
Tfi DNA ligase is not essential for the enzyme activity. The enzymatic properties of
Tfi-M1 mutant (deleted the BRCT domain) were slightly different from those of wild-type and the nick-closing activity of
Tfi-M1 mutant was approximately 50% compared with that of wild-type.</abstract><cop>Oxford</cop><pub>Elsevier B.V</pub><pmid>15268945</pmid><doi>10.1016/j.femsle.2004.06.018</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0378-1097 |
| ispartof | FEMS microbiology letters, 2004-08, Vol.237 (1), p.111-118 |
| issn | 0378-1097 1574-6968 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_66736589 |
| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current); Wiley Online Library All Journals; Alma/SFX Local Collection |
| subjects | Adenosine Monophosphate - metabolism Bacteriology Biological and medical sciences Cloning, Molecular Coenzymes - pharmacology DNA binding activity DNA Ligases - genetics DNA Ligases - isolation & purification DNA Ligases - metabolism DNA Mutational Analysis DNA-Binding Proteins - genetics DNA-Binding Proteins - physiology Enzyme Stability Escherichia coli - genetics Escherichia coli - growth & development Escherichia coli - metabolism Fundamental and applied biological sciences. Psychology Genetic Complementation Test Helix-Turn-Helix Motifs - genetics Helix-Turn-Helix Motifs - physiology Hydrogen-Ion Concentration Microbiology Miscellaneous Nick-closing activity Protein Structure, Tertiary Recombinant Proteins - genetics Recombinant Proteins - isolation & purification Recombinant Proteins - metabolism Self-adenylation Sequence Deletion Temperature Thermus - enzymology Thermus filiformis ( Tfi) DNA ligase Time Factors Zinc Fingers - genetics Zinc Fingers - physiology |
| title | Mutational analyses of the thermostable NAD +-dependent DNA ligase from Thermus filiformis |
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