Purification and initial characterization of AhrC: the regulator of arginine metabolism genes in Bacillus subtilis
Summary The arginine‐dependent repressor‐activator from Bacillus subtilis, AhrC, has been overexpressed in Escherichia coli and purified to homogeneity. AhrC, expressed in E. coli, is able to repress a Bacillus promoter (argCp), which lies upstream of the argC gene. The purified protein is a hexamer...
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| Veröffentlicht in: | Molecular microbiology 1992-01, Vol.6 (2), p.267-275 |
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| creator | Czaplewski, L. G. North, A. K. Smith, M. C. M. Baumberg, S. Stockley, P. G. |
| description | Summary
The arginine‐dependent repressor‐activator from Bacillus subtilis, AhrC, has been overexpressed in Escherichia coli and purified to homogeneity. AhrC, expressed in E. coli, is able to repress a Bacillus promoter (argCp), which lies upstream of the argC gene. The purified protein is a hexamer with a subunit molecular mass of 16.7 kDa. Its ability to recognize DNA has been examined in vitro using argCp in both DNase I and hydroxyl radical protection assays. AhrC binds at two distinct sites within the argCp fragment. One site, argCO1, with the highest affinity for protein, is located within the 5′ promoter sequences, whilst the other, argCO2, is within the coding region of argC. The data are consistent with the binding of a single hexamer of AhrC to argCO1 via four of its subunits, possibly allowing the remaining two subunits to bind at argCO2in vivo forming a repression loop similar to those observed for the E coli Lac repressor. |
| doi_str_mv | 10.1111/j.1365-2958.1992.tb02008.x |
| format | Article |
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The arginine‐dependent repressor‐activator from Bacillus subtilis, AhrC, has been overexpressed in Escherichia coli and purified to homogeneity. AhrC, expressed in E. coli, is able to repress a Bacillus promoter (argCp), which lies upstream of the argC gene. The purified protein is a hexamer with a subunit molecular mass of 16.7 kDa. Its ability to recognize DNA has been examined in vitro using argCp in both DNase I and hydroxyl radical protection assays. AhrC binds at two distinct sites within the argCp fragment. One site, argCO1, with the highest affinity for protein, is located within the 5′ promoter sequences, whilst the other, argCO2, is within the coding region of argC. The data are consistent with the binding of a single hexamer of AhrC to argCO1 via four of its subunits, possibly allowing the remaining two subunits to bind at argCO2in vivo forming a repression loop similar to those observed for the E coli Lac repressor.</description><identifier>ISSN: 0950-382X</identifier><identifier>EISSN: 1365-2958</identifier><identifier>DOI: 10.1111/j.1365-2958.1992.tb02008.x</identifier><identifier>PMID: 1312212</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Amino Acid Sequence ; Arginine - metabolism ; Bacillus subtilis ; Bacillus subtilis - genetics ; Bacillus subtilis - metabolism ; Bacterial Proteins ; Base Sequence ; Biological and medical sciences ; Cyclic AMP Receptor Protein - chemistry ; Cyclic AMP Receptor Protein - genetics ; Cyclic AMP Receptor Protein - isolation & purification ; Cyclic AMP Receptor Protein - metabolism ; DNA, Bacterial - metabolism ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Escherichia coli Proteins ; Fundamental and applied biological sciences. Psychology ; Genes, Bacterial ; Microbiology ; Molecular Sequence Data ; Molecular Weight ; Promoter Regions, Genetic ; Repressor Proteins - chemistry ; Repressor Proteins - genetics ; Repressor Proteins - isolation & purification ; Repressor Proteins - metabolism ; Techniques used in virology ; Virology</subject><ispartof>Molecular microbiology, 1992-01, Vol.6 (2), p.267-275</ispartof><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4957-b5edcdd543082139e60a2f156b35f2caef6c3b03438d5b91164655bb7433f6da3</citedby><cites>FETCH-LOGICAL-c4957-b5edcdd543082139e60a2f156b35f2caef6c3b03438d5b91164655bb7433f6da3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1365-2958.1992.tb02008.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1365-2958.1992.tb02008.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,4024,27923,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5050789$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1312212$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Czaplewski, L. G.</creatorcontrib><creatorcontrib>North, A. K.</creatorcontrib><creatorcontrib>Smith, M. C. M.</creatorcontrib><creatorcontrib>Baumberg, S.</creatorcontrib><creatorcontrib>Stockley, P. G.</creatorcontrib><title>Purification and initial characterization of AhrC: the regulator of arginine metabolism genes in Bacillus subtilis</title><title>Molecular microbiology</title><addtitle>Mol Microbiol</addtitle><description>Summary
The arginine‐dependent repressor‐activator from Bacillus subtilis, AhrC, has been overexpressed in Escherichia coli and purified to homogeneity. AhrC, expressed in E. coli, is able to repress a Bacillus promoter (argCp), which lies upstream of the argC gene. The purified protein is a hexamer with a subunit molecular mass of 16.7 kDa. Its ability to recognize DNA has been examined in vitro using argCp in both DNase I and hydroxyl radical protection assays. AhrC binds at two distinct sites within the argCp fragment. One site, argCO1, with the highest affinity for protein, is located within the 5′ promoter sequences, whilst the other, argCO2, is within the coding region of argC. The data are consistent with the binding of a single hexamer of AhrC to argCO1 via four of its subunits, possibly allowing the remaining two subunits to bind at argCO2in vivo forming a repression loop similar to those observed for the E coli Lac repressor.</description><subject>Amino Acid Sequence</subject><subject>Arginine - metabolism</subject><subject>Bacillus subtilis</subject><subject>Bacillus subtilis - genetics</subject><subject>Bacillus subtilis - metabolism</subject><subject>Bacterial Proteins</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cyclic AMP Receptor Protein - chemistry</subject><subject>Cyclic AMP Receptor Protein - genetics</subject><subject>Cyclic AMP Receptor Protein - isolation & purification</subject><subject>Cyclic AMP Receptor Protein - metabolism</subject><subject>DNA, Bacterial - metabolism</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>Genes, Bacterial</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Molecular Weight</subject><subject>Promoter Regions, Genetic</subject><subject>Repressor Proteins - chemistry</subject><subject>Repressor Proteins - genetics</subject><subject>Repressor Proteins - isolation & purification</subject><subject>Repressor Proteins - metabolism</subject><subject>Techniques used in virology</subject><subject>Virology</subject><issn>0950-382X</issn><issn>1365-2958</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkU1r3DAQhkVpSLdpf0JBlNKbXX1YspVDIV36EUhIDy30JkayvKtFtlNJpkl_fWy8pMeSuQjmfd7RMC9Cbykp6VwfDiXlUhRMiaakSrEyG8IIacq7Z2jzKD1HG6IEKXjDfr1AL1M6EEI5kfwUnVJOGaNsg-L3KfrOW8h-HDAMLfaDzx4CtnuIYLOL_u8qjh2-2MftOc57h6PbTQHyGJc2xN3sGhzuXQYzBp96vHODS_Mw_AmsD2FKOE0m-1l7hU46CMm9Pr5n6OeXzz-234qrm6-X24urwlZK1IURrrVtKypOGka5cpIA66iQhouOWXCdtNwQXvGmFUZRKisphDF1xXknW-Bn6P069zaOvyeXsu59si4EGNw4JV2zhquqVv8FqaQzSskMnq-gjWNK0XX6Nvoe4r2mRC_J6INezq-X8-slGX1MRt_N5jfHXybTu_afdY1i1t8ddUgWQhdhsD49YoIIUjfLsh9X7I8P7v4JC-jr60sma_4AQ0esGg</recordid><startdate>199201</startdate><enddate>199201</enddate><creator>Czaplewski, L. G.</creator><creator>North, A. K.</creator><creator>Smith, M. C. M.</creator><creator>Baumberg, S.</creator><creator>Stockley, P. G.</creator><general>Blackwell Publishing Ltd</general><general>Blackwell Science</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>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>199201</creationdate><title>Purification and initial characterization of AhrC: the regulator of arginine metabolism genes in Bacillus subtilis</title><author>Czaplewski, L. G. ; North, A. K. ; Smith, M. C. M. ; Baumberg, S. ; Stockley, P. G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4957-b5edcdd543082139e60a2f156b35f2caef6c3b03438d5b91164655bb7433f6da3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Amino Acid Sequence</topic><topic>Arginine - metabolism</topic><topic>Bacillus subtilis</topic><topic>Bacillus subtilis - genetics</topic><topic>Bacillus subtilis - metabolism</topic><topic>Bacterial Proteins</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cyclic AMP Receptor Protein - chemistry</topic><topic>Cyclic AMP Receptor Protein - genetics</topic><topic>Cyclic AMP Receptor Protein - isolation & purification</topic><topic>Cyclic AMP Receptor Protein - metabolism</topic><topic>DNA, Bacterial - metabolism</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>Genes, Bacterial</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>Promoter Regions, Genetic</topic><topic>Repressor Proteins - chemistry</topic><topic>Repressor Proteins - genetics</topic><topic>Repressor Proteins - isolation & purification</topic><topic>Repressor Proteins - metabolism</topic><topic>Techniques used in virology</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Czaplewski, L. G.</creatorcontrib><creatorcontrib>North, A. K.</creatorcontrib><creatorcontrib>Smith, M. C. M.</creatorcontrib><creatorcontrib>Baumberg, S.</creatorcontrib><creatorcontrib>Stockley, P. G.</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>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Czaplewski, L. G.</au><au>North, A. K.</au><au>Smith, M. C. M.</au><au>Baumberg, S.</au><au>Stockley, P. G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification and initial characterization of AhrC: the regulator of arginine metabolism genes in Bacillus subtilis</atitle><jtitle>Molecular microbiology</jtitle><addtitle>Mol Microbiol</addtitle><date>1992-01</date><risdate>1992</risdate><volume>6</volume><issue>2</issue><spage>267</spage><epage>275</epage><pages>267-275</pages><issn>0950-382X</issn><eissn>1365-2958</eissn><abstract>Summary
The arginine‐dependent repressor‐activator from Bacillus subtilis, AhrC, has been overexpressed in Escherichia coli and purified to homogeneity. AhrC, expressed in E. coli, is able to repress a Bacillus promoter (argCp), which lies upstream of the argC gene. The purified protein is a hexamer with a subunit molecular mass of 16.7 kDa. Its ability to recognize DNA has been examined in vitro using argCp in both DNase I and hydroxyl radical protection assays. AhrC binds at two distinct sites within the argCp fragment. One site, argCO1, with the highest affinity for protein, is located within the 5′ promoter sequences, whilst the other, argCO2, is within the coding region of argC. The data are consistent with the binding of a single hexamer of AhrC to argCO1 via four of its subunits, possibly allowing the remaining two subunits to bind at argCO2in vivo forming a repression loop similar to those observed for the E coli Lac repressor.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>1312212</pmid><doi>10.1111/j.1365-2958.1992.tb02008.x</doi><tpages>9</tpages></addata></record> |
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| ispartof | Molecular microbiology, 1992-01, Vol.6 (2), p.267-275 |
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| subjects | Amino Acid Sequence Arginine - metabolism Bacillus subtilis Bacillus subtilis - genetics Bacillus subtilis - metabolism Bacterial Proteins Base Sequence Biological and medical sciences Cyclic AMP Receptor Protein - chemistry Cyclic AMP Receptor Protein - genetics Cyclic AMP Receptor Protein - isolation & purification Cyclic AMP Receptor Protein - metabolism DNA, Bacterial - metabolism Escherichia coli - genetics Escherichia coli - metabolism Escherichia coli Proteins Fundamental and applied biological sciences. Psychology Genes, Bacterial Microbiology Molecular Sequence Data Molecular Weight Promoter Regions, Genetic Repressor Proteins - chemistry Repressor Proteins - genetics Repressor Proteins - isolation & purification Repressor Proteins - metabolism Techniques used in virology Virology |
| title | Purification and initial characterization of AhrC: the regulator of arginine metabolism genes in Bacillus subtilis |
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