GcvA binding site 1 in the gcvTHP promoter of Escherichia coli is required for GcvA-mediated repression but not for GcvA-mediated activation
Department of Microbiology, The University of Iowa, Iowa City, IA 52242, USA 1 Author for correspondence: George V. Stauffer. Tel: +1 319 335 7791. Fax: +1 319 335 9006. e-mail: george-stauffer{at}uiowa.edu GcvA binds to three sites in the gcvTHP control region, from base -34 to -69 (site 1), from b...
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| Veröffentlicht in: | Microbiology (Society for General Microbiology) 2000-11, Vol.146 (11), p.2909-2918 |
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| creator | Wonderling, Laura D Urbanowski, Mark L Stauffer, George V |
| description | Department of Microbiology, The University of Iowa, Iowa City, IA 52242, USA 1
Author for correspondence: George V. Stauffer. Tel: +1 319 335 7791. Fax: +1 319 335 9006. e-mail: george-stauffer{at}uiowa.edu
GcvA binds to three sites in the gcvTHP control region, from base -34 to -69 (site 1), from base -214 to -241 (site 2) and from base -242 to -271 (site 3). Previous results suggested that sites 3 and 2 are required for both GcvA-dependent activation and repression of a gcvT :: lacZ fusion. However, the results were less clear as to the role of site 1. To determine the role of site 1 in regulation, single and multiple base changes were made in site 1 and tested for their ability to alter GcvA-mediated activation and GcvA/GcvR-mediated repression. Several of the mutants were also tested for effects on GcvA binding to site 1 and the ability of GcvA to bend DNA at site 1. The results are consistent with site 1 playing primarily a role in negative regulation of the gcvTHP operon.
Keywords: gcvTHP , gcvT :: lacZ , GcvA, GcvR, repression Abbreviations: CRP, cyclic AMP receptor protein; GCV, glycine cleavage; RNAP, RNA polymerase; WT, wild-type |
| doi_str_mv | 10.1099/00221287-146-11-2909 |
| format | Article |
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Author for correspondence: George V. Stauffer. Tel: +1 319 335 7791. Fax: +1 319 335 9006. e-mail: george-stauffer{at}uiowa.edu
GcvA binds to three sites in the gcvTHP control region, from base -34 to -69 (site 1), from base -214 to -241 (site 2) and from base -242 to -271 (site 3). Previous results suggested that sites 3 and 2 are required for both GcvA-dependent activation and repression of a gcvT :: lacZ fusion. However, the results were less clear as to the role of site 1. To determine the role of site 1 in regulation, single and multiple base changes were made in site 1 and tested for their ability to alter GcvA-mediated activation and GcvA/GcvR-mediated repression. Several of the mutants were also tested for effects on GcvA binding to site 1 and the ability of GcvA to bend DNA at site 1. The results are consistent with site 1 playing primarily a role in negative regulation of the gcvTHP operon.
Keywords: gcvTHP , gcvT :: lacZ , GcvA, GcvR, repression Abbreviations: CRP, cyclic AMP receptor protein; GCV, glycine cleavage; RNAP, RNA polymerase; WT, wild-type</description><identifier>ISSN: 1350-0872</identifier><identifier>EISSN: 1465-2080</identifier><identifier>DOI: 10.1099/00221287-146-11-2909</identifier><identifier>PMID: 11065369</identifier><language>eng</language><publisher>Reading: Soc General Microbiol</publisher><subject>Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Bacteriology ; Binding Sites - genetics ; Biological and medical sciences ; Conserved Sequence ; DNA, Bacterial - chemistry ; DNA, Bacterial - genetics ; DNA, Bacterial - metabolism ; DNA-Binding Proteins - genetics ; DNA-Binding Proteins - metabolism ; Escherichia coli ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Escherichia coli Proteins ; Fundamental and applied biological sciences. Psychology ; GcvA protein ; gcvH gene ; gcvP gene ; GcvR protein ; gcvT gene ; Gene Expression Regulation, Bacterial ; Genes, Bacterial ; Genetics ; Microbiology ; Nucleic Acid Conformation ; Operon ; Phenotype ; Point Mutation ; Promoter Regions, Genetic ; Transcription Factors - genetics ; Transcription Factors - metabolism</subject><ispartof>Microbiology (Society for General Microbiology), 2000-11, Vol.146 (11), p.2909-2918</ispartof><rights>2001 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c443t-532354fa76e8b4e6e63fa65b8cbad8294582c79b490c90a8c64b233d37e80af73</citedby><cites>FETCH-LOGICAL-c443t-532354fa76e8b4e6e63fa65b8cbad8294582c79b490c90a8c64b233d37e80af73</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,777,781,27905,27906</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=827439$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11065369$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wonderling, Laura D</creatorcontrib><creatorcontrib>Urbanowski, Mark L</creatorcontrib><creatorcontrib>Stauffer, George V</creatorcontrib><title>GcvA binding site 1 in the gcvTHP promoter of Escherichia coli is required for GcvA-mediated repression but not for GcvA-mediated activation</title><title>Microbiology (Society for General Microbiology)</title><addtitle>Microbiology</addtitle><description>Department of Microbiology, The University of Iowa, Iowa City, IA 52242, USA 1
Author for correspondence: George V. Stauffer. Tel: +1 319 335 7791. Fax: +1 319 335 9006. e-mail: george-stauffer{at}uiowa.edu
GcvA binds to three sites in the gcvTHP control region, from base -34 to -69 (site 1), from base -214 to -241 (site 2) and from base -242 to -271 (site 3). Previous results suggested that sites 3 and 2 are required for both GcvA-dependent activation and repression of a gcvT :: lacZ fusion. However, the results were less clear as to the role of site 1. To determine the role of site 1 in regulation, single and multiple base changes were made in site 1 and tested for their ability to alter GcvA-mediated activation and GcvA/GcvR-mediated repression. Several of the mutants were also tested for effects on GcvA binding to site 1 and the ability of GcvA to bend DNA at site 1. The results are consistent with site 1 playing primarily a role in negative regulation of the gcvTHP operon.
Keywords: gcvTHP , gcvT :: lacZ , GcvA, GcvR, repression Abbreviations: CRP, cyclic AMP receptor protein; GCV, glycine cleavage; RNAP, RNA polymerase; WT, wild-type</description><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Bacteriology</subject><subject>Binding Sites - genetics</subject><subject>Biological and medical sciences</subject><subject>Conserved Sequence</subject><subject>DNA, Bacterial - chemistry</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA, Bacterial - metabolism</subject><subject>DNA-Binding Proteins - genetics</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Escherichia coli Proteins</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>GcvA protein</subject><subject>gcvH gene</subject><subject>gcvP gene</subject><subject>GcvR protein</subject><subject>gcvT gene</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Genes, Bacterial</subject><subject>Genetics</subject><subject>Microbiology</subject><subject>Nucleic Acid Conformation</subject><subject>Operon</subject><subject>Phenotype</subject><subject>Point Mutation</subject><subject>Promoter Regions, Genetic</subject><subject>Transcription Factors - genetics</subject><subject>Transcription Factors - metabolism</subject><issn>1350-0872</issn><issn>1465-2080</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcuKFDEUhgtxcC76BiIBwcVAxtyqkiyHYZwRBnQxrkOSOtUVqar0JKkW38GHNk23l4XgKrfv_DnJ1zSvKbmiROv3hDBGmZKYig5Tipkm-llzVlctZkSR53XOW4KJkuy0Oc_5KyH1kNAXzSmlpGt5p8-aH3d-d41cWPqwbFAOBRBFYUFlBLTxu8f7z2ib4hwLJBQHdJv9CCn4MVjk4xRQyCjB0xoS9GiICe3j8Ax9sKXuJNgmyDnEBbm1oCWWf0DWl7CzpUIvm5PBThleHceL5suH28ebe_zw6e7jzfUD9kLwglvOeCsGKztQTkAHHR9s1zrlne0V06JVzEvthCZeE6t8JxzjvOcSFLGD5BfNu0NufdrTCrmYOWQP02QXiGs2knEtGaX_BalUopK8guIA-hRzTjCYbQqzTd8NJWavy_zSZaoDQ6nZ66plb475q6v_8afo6KcCb4-Azd5OQ7KLD_k3p5gUfE9dHqgxbMZv1YXZwDKH2osLsbbs_77zJ1toq2I</recordid><startdate>20001101</startdate><enddate>20001101</enddate><creator>Wonderling, Laura D</creator><creator>Urbanowski, Mark L</creator><creator>Stauffer, George V</creator><general>Soc General Microbiol</general><general>Society for General Microbiology</general><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>20001101</creationdate><title>GcvA binding site 1 in the gcvTHP promoter of Escherichia coli is required for GcvA-mediated repression but not for GcvA-mediated activation</title><author>Wonderling, Laura D ; Urbanowski, Mark L ; Stauffer, George V</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c443t-532354fa76e8b4e6e63fa65b8cbad8294582c79b490c90a8c64b233d37e80af73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Bacteriology</topic><topic>Binding Sites - genetics</topic><topic>Biological and medical sciences</topic><topic>Conserved Sequence</topic><topic>DNA, Bacterial - chemistry</topic><topic>DNA, Bacterial - genetics</topic><topic>DNA, Bacterial - metabolism</topic><topic>DNA-Binding Proteins - genetics</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>Escherichia coli Proteins</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>GcvA protein</topic><topic>gcvH gene</topic><topic>gcvP gene</topic><topic>GcvR protein</topic><topic>gcvT gene</topic><topic>Gene Expression Regulation, Bacterial</topic><topic>Genes, Bacterial</topic><topic>Genetics</topic><topic>Microbiology</topic><topic>Nucleic Acid Conformation</topic><topic>Operon</topic><topic>Phenotype</topic><topic>Point Mutation</topic><topic>Promoter Regions, Genetic</topic><topic>Transcription Factors - genetics</topic><topic>Transcription Factors - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wonderling, Laura D</creatorcontrib><creatorcontrib>Urbanowski, Mark L</creatorcontrib><creatorcontrib>Stauffer, George V</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>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>Microbiology (Society for General Microbiology)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wonderling, Laura D</au><au>Urbanowski, Mark L</au><au>Stauffer, George V</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>GcvA binding site 1 in the gcvTHP promoter of Escherichia coli is required for GcvA-mediated repression but not for GcvA-mediated activation</atitle><jtitle>Microbiology (Society for General Microbiology)</jtitle><addtitle>Microbiology</addtitle><date>2000-11-01</date><risdate>2000</risdate><volume>146</volume><issue>11</issue><spage>2909</spage><epage>2918</epage><pages>2909-2918</pages><issn>1350-0872</issn><eissn>1465-2080</eissn><abstract>Department of Microbiology, The University of Iowa, Iowa City, IA 52242, USA 1
Author for correspondence: George V. Stauffer. Tel: +1 319 335 7791. Fax: +1 319 335 9006. e-mail: george-stauffer{at}uiowa.edu
GcvA binds to three sites in the gcvTHP control region, from base -34 to -69 (site 1), from base -214 to -241 (site 2) and from base -242 to -271 (site 3). Previous results suggested that sites 3 and 2 are required for both GcvA-dependent activation and repression of a gcvT :: lacZ fusion. However, the results were less clear as to the role of site 1. To determine the role of site 1 in regulation, single and multiple base changes were made in site 1 and tested for their ability to alter GcvA-mediated activation and GcvA/GcvR-mediated repression. Several of the mutants were also tested for effects on GcvA binding to site 1 and the ability of GcvA to bend DNA at site 1. The results are consistent with site 1 playing primarily a role in negative regulation of the gcvTHP operon.
Keywords: gcvTHP , gcvT :: lacZ , GcvA, GcvR, repression Abbreviations: CRP, cyclic AMP receptor protein; GCV, glycine cleavage; RNAP, RNA polymerase; WT, wild-type</abstract><cop>Reading</cop><pub>Soc General Microbiol</pub><pmid>11065369</pmid><doi>10.1099/00221287-146-11-2909</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Bacterial Proteins - genetics Bacterial Proteins - metabolism Bacteriology Binding Sites - genetics Biological and medical sciences Conserved Sequence DNA, Bacterial - chemistry DNA, Bacterial - genetics DNA, Bacterial - metabolism DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism Escherichia coli Escherichia coli - genetics Escherichia coli - metabolism Escherichia coli Proteins Fundamental and applied biological sciences. Psychology GcvA protein gcvH gene gcvP gene GcvR protein gcvT gene Gene Expression Regulation, Bacterial Genes, Bacterial Genetics Microbiology Nucleic Acid Conformation Operon Phenotype Point Mutation Promoter Regions, Genetic Transcription Factors - genetics Transcription Factors - metabolism |
| title | GcvA binding site 1 in the gcvTHP promoter of Escherichia coli is required for GcvA-mediated repression but not for GcvA-mediated activation |
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