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Induced Bending of Plasmid pLS1 DNA by the Plasmid-encoded Protein RepA
The broad host range streptococcal plasmid pLS1 encodes for a 5.1-kDa repressor protein, RepA. This protein has affinity for DNA (linear or supercoiled) and is translated from the same mRNA as the replication initiator protein RepB. By gel retardation assays, we observed that RepA shows specificity...
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| Published in: | The Journal of biological chemistry 1989-12, Vol.264 (35), p.21334-21339 |
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| Main Authors: | , , , , , |
| Format: | Article |
| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c496t-34919782299812efad0d614a60e9ec5741148901786f0cfbc787f1636500db763 |
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| cites | cdi_FETCH-LOGICAL-c496t-34919782299812efad0d614a60e9ec5741148901786f0cfbc787f1636500db763 |
| container_end_page | 21339 |
| container_issue | 35 |
| container_start_page | 21334 |
| container_title | The Journal of biological chemistry |
| container_volume | 264 |
| creator | Pérez-Martín, J del Solar, G H Lurz, R de la Campa, A G Dobrinski, B Espinosa, M |
| description | The broad host range streptococcal plasmid pLS1 encodes for a 5.1-kDa repressor protein, RepA. This protein has affinity for DNA (linear or supercoiled) and is translated from the same mRNA as the replication initiator protein RepB. By gel retardation assays, we observed that RepA shows specificity for binding to the plasmid HinfID fragment, which includes the target of the protein. The target of RepA within the plasmid DNA molecule has been located around the plasmid single site ApaLI. This site is included in a region that contains the promoter for the repA and repB genes and is contiguous to the plasmid ori(+). A complex sequence-directed DNA curvature is observed in this region of pLS1. Upon addition of RepA to plasmid linear DNA or to circularly permuted restriction fragments, this intrinsic curvature was greatly enhanced. Thus, a strong RepA-induced bending could be located in the vicinity of the ApaLI site. Visualization of the bent DNA was achieved by electron microscopy of complexes between RepA and plasmid DNA fragments containing the RepA target. |
| doi_str_mv | 10.1016/S0021-9258(19)30084-5 |
| format | article |
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This protein has affinity for DNA (linear or supercoiled) and is translated from the same mRNA as the replication initiator protein RepB. By gel retardation assays, we observed that RepA shows specificity for binding to the plasmid HinfID fragment, which includes the target of the protein. The target of RepA within the plasmid DNA molecule has been located around the plasmid single site ApaLI. This site is included in a region that contains the promoter for the repA and repB genes and is contiguous to the plasmid ori(+). A complex sequence-directed DNA curvature is observed in this region of pLS1. Upon addition of RepA to plasmid linear DNA or to circularly permuted restriction fragments, this intrinsic curvature was greatly enhanced. Thus, a strong RepA-induced bending could be located in the vicinity of the ApaLI site. Visualization of the bent DNA was achieved by electron microscopy of complexes between RepA and plasmid DNA fragments containing the RepA target.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/S0021-9258(19)30084-5</identifier><identifier>PMID: 2592378</identifier><identifier>CODEN: JBCHA3</identifier><language>eng</language><publisher>Bethesda, MD: Elsevier Inc</publisher><subject>Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Base Sequence ; Biological and medical sciences ; Cloning, Molecular - methods ; Computer Simulation ; DNA Helicases ; DNA Replication ; DNA, Bacterial - genetics ; DNA, Bacterial - metabolism ; DNA, Bacterial - ultrastructure ; DNA-Binding Proteins ; Fundamental and applied biological sciences. Psychology ; Interactions. Associations ; Intermolecular phenomena ; Microscopy, Electron ; Molecular biophysics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Plasmids ; Proteins ; Restriction Mapping ; Streptococcus pneumoniae - genetics ; Substrate Specificity ; Trans-Activators</subject><ispartof>The Journal of biological chemistry, 1989-12, Vol.264 (35), p.21334-21339</ispartof><rights>1989 © 1989 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><rights>1990 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c496t-34919782299812efad0d614a60e9ec5741148901786f0cfbc787f1636500db763</citedby><cites>FETCH-LOGICAL-c496t-34919782299812efad0d614a60e9ec5741148901786f0cfbc787f1636500db763</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,786,790,27957,27958</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6692089$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2592378$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pérez-Martín, J</creatorcontrib><creatorcontrib>del Solar, G H</creatorcontrib><creatorcontrib>Lurz, R</creatorcontrib><creatorcontrib>de la Campa, A G</creatorcontrib><creatorcontrib>Dobrinski, B</creatorcontrib><creatorcontrib>Espinosa, M</creatorcontrib><title>Induced Bending of Plasmid pLS1 DNA by the Plasmid-encoded Protein RepA</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The broad host range streptococcal plasmid pLS1 encodes for a 5.1-kDa repressor protein, RepA. This protein has affinity for DNA (linear or supercoiled) and is translated from the same mRNA as the replication initiator protein RepB. By gel retardation assays, we observed that RepA shows specificity for binding to the plasmid HinfID fragment, which includes the target of the protein. The target of RepA within the plasmid DNA molecule has been located around the plasmid single site ApaLI. This site is included in a region that contains the promoter for the repA and repB genes and is contiguous to the plasmid ori(+). A complex sequence-directed DNA curvature is observed in this region of pLS1. Upon addition of RepA to plasmid linear DNA or to circularly permuted restriction fragments, this intrinsic curvature was greatly enhanced. Thus, a strong RepA-induced bending could be located in the vicinity of the ApaLI site. Visualization of the bent DNA was achieved by electron microscopy of complexes between RepA and plasmid DNA fragments containing the RepA target.</description><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cloning, Molecular - methods</subject><subject>Computer Simulation</subject><subject>DNA Helicases</subject><subject>DNA Replication</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA, Bacterial - metabolism</subject><subject>DNA, Bacterial - ultrastructure</subject><subject>DNA-Binding Proteins</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Interactions. Associations</subject><subject>Intermolecular phenomena</subject><subject>Microscopy, Electron</subject><subject>Molecular biophysics</subject><subject>Molecular Sequence Data</subject><subject>Nucleic Acid Conformation</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Restriction Mapping</subject><subject>Streptococcus pneumoniae - genetics</subject><subject>Substrate Specificity</subject><subject>Trans-Activators</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><recordid>eNqFkV9vFCEUxYnRtGv1IzSZh8bow1guDAw8NWvV2mRTG2uTvhEG7nRp5s8Ks5p-e9nuuj6WFxLu73AOB0KOgX4ECvL0hlIGpWZCvQf9gVOqqlK8IDOgipdcwN1LMtsjh-R1Sg80r0rDATlgQjNeqxm5uBz82qEvPuHgw3BfjG1x3dnUB1-sFjdQfL6aF81jMS3x33mJgxt9llzHccIwFD9wNX9DXrW2S_h2tx-R269ffp5_KxffLy7P54vSVVpOJc_2ulaMaa2AYWs99RIqKylqdKKuACqlKdRKttS1jatV3YLkUlDqm1ryI_Jue-8qjr_WmCbTh-Sw6-yA4zqZWnMp89ueBUFUUmbrDIot6OKYUsTWrGLobXw0QM2mafPUtNnUaECbp6aNyLrjncG66dHvVbtq8_xkN7fJ2a6NdnAh7bFNSqr0f2wZ7pd_QkTThNEtsTdMVoYLw4DzTcqzLYa53N8Bo0ku5I9AnyVuMn4Mz-T9Cz3Cof0</recordid><startdate>19891215</startdate><enddate>19891215</enddate><creator>Pérez-Martín, J</creator><creator>del Solar, G H</creator><creator>Lurz, R</creator><creator>de la Campa, A G</creator><creator>Dobrinski, B</creator><creator>Espinosa, M</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>IQODW</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>7QL</scope><scope>7TM</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>19891215</creationdate><title>Induced Bending of Plasmid pLS1 DNA by the Plasmid-encoded Protein RepA</title><author>Pérez-Martín, J ; del Solar, G H ; Lurz, R ; de la Campa, A G ; Dobrinski, B ; Espinosa, M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c496t-34919782299812efad0d614a60e9ec5741148901786f0cfbc787f1636500db763</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cloning, Molecular - methods</topic><topic>Computer Simulation</topic><topic>DNA Helicases</topic><topic>DNA Replication</topic><topic>DNA, Bacterial - genetics</topic><topic>DNA, Bacterial - metabolism</topic><topic>DNA, Bacterial - ultrastructure</topic><topic>DNA-Binding Proteins</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Interactions. Associations</topic><topic>Intermolecular phenomena</topic><topic>Microscopy, Electron</topic><topic>Molecular biophysics</topic><topic>Molecular Sequence Data</topic><topic>Nucleic Acid Conformation</topic><topic>Plasmids</topic><topic>Proteins</topic><topic>Restriction Mapping</topic><topic>Streptococcus pneumoniae - genetics</topic><topic>Substrate Specificity</topic><topic>Trans-Activators</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pérez-Martín, J</creatorcontrib><creatorcontrib>del Solar, G H</creatorcontrib><creatorcontrib>Lurz, R</creatorcontrib><creatorcontrib>de la Campa, A G</creatorcontrib><creatorcontrib>Dobrinski, B</creatorcontrib><creatorcontrib>Espinosa, M</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Environmental Sciences and Pollution Management</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>Pérez-Martín, J</au><au>del Solar, G H</au><au>Lurz, R</au><au>de la Campa, A G</au><au>Dobrinski, B</au><au>Espinosa, M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Induced Bending of Plasmid pLS1 DNA by the Plasmid-encoded Protein RepA</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1989-12-15</date><risdate>1989</risdate><volume>264</volume><issue>35</issue><spage>21334</spage><epage>21339</epage><pages>21334-21339</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><notes>ObjectType-Article-2</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-1</notes><notes>content type line 23</notes><notes>ObjectType-Article-1</notes><notes>ObjectType-Feature-2</notes><abstract>The broad host range streptococcal plasmid pLS1 encodes for a 5.1-kDa repressor protein, RepA. This protein has affinity for DNA (linear or supercoiled) and is translated from the same mRNA as the replication initiator protein RepB. By gel retardation assays, we observed that RepA shows specificity for binding to the plasmid HinfID fragment, which includes the target of the protein. The target of RepA within the plasmid DNA molecule has been located around the plasmid single site ApaLI. This site is included in a region that contains the promoter for the repA and repB genes and is contiguous to the plasmid ori(+). A complex sequence-directed DNA curvature is observed in this region of pLS1. Upon addition of RepA to plasmid linear DNA or to circularly permuted restriction fragments, this intrinsic curvature was greatly enhanced. Thus, a strong RepA-induced bending could be located in the vicinity of the ApaLI site. Visualization of the bent DNA was achieved by electron microscopy of complexes between RepA and plasmid DNA fragments containing the RepA target.</abstract><cop>Bethesda, MD</cop><pub>Elsevier Inc</pub><pmid>2592378</pmid><doi>10.1016/S0021-9258(19)30084-5</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1989-12, Vol.264 (35), p.21334-21339 |
| issn | 0021-9258 1083-351X |
| language | eng |
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| source | BACON - Elsevier - GLOBAL_SCIENCEDIRECT-OPENACCESS |
| subjects | Bacterial Proteins - genetics Bacterial Proteins - metabolism Base Sequence Biological and medical sciences Cloning, Molecular - methods Computer Simulation DNA Helicases DNA Replication DNA, Bacterial - genetics DNA, Bacterial - metabolism DNA, Bacterial - ultrastructure DNA-Binding Proteins Fundamental and applied biological sciences. Psychology Interactions. Associations Intermolecular phenomena Microscopy, Electron Molecular biophysics Molecular Sequence Data Nucleic Acid Conformation Plasmids Proteins Restriction Mapping Streptococcus pneumoniae - genetics Substrate Specificity Trans-Activators |
| title | Induced Bending of Plasmid pLS1 DNA by the Plasmid-encoded Protein RepA |
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