Transcription-Induced Barriers to Supercoil Diffusion in the Salmonella typhimurium Chromosome
Transcription and replication both influence and are influenced by superhelical changes in DNA. Explaining how supercoil movement is channeled in living chromosomes has been a major problem for 30 years. Transcription of membrane-associated proteins leads to localized hypersupercoiling of plasmid DN...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2004-03, Vol.101 (10), p.3398-3403 |
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| creator | Deng, Shuang Stein, Richard A. Higgins, N. Patrick Gottesman, Susan |
| description | Transcription and replication both influence and are influenced by superhelical changes in DNA. Explaining how supercoil movement is channeled in living chromosomes has been a major problem for 30 years. Transcription of membrane-associated proteins leads to localized hypersupercoiling of plasmid DNA, and this behavior indicates the presence of aberrant supercoil diffusion. Using the lambda Red recombination system, we constructed model domains in the Salmonella typhimurium chromosome to analyze supercoiling dynamics of regions encoding membrane proteins. Regulation of Tn10-derived tetracycline resistance involves a repressor, TetR, and a membrane-bound export pump, TetA. Strains deficient in TetR activity had 60-fold higher transcription levels (from PA) than TetR-positive strains. High tetA transcription caused a 10- to 80-fold decrease in the γδ resolution efficiency for the domain that includes the Tet module. Replacing tetA with genes encoding cytosolic proteins LacZ and Kan also caused the appearance of supercoil diffusion barriers in a defined region of the chromosome. In strains containing a functional TetR located next to a regulated lacZ reporter (PRtetR-PAlacZ), induction of transcription with chlortetracycline caused a 5-fold drop in resolution efficiency in the test domain interval. A short half-life resolves showed that barriers appeared and disappeared over a 10- to 20-min span. These studies demonstrate the importance of transcription in chromosome structure and the plasticity of supercoil domains in bacterial chromosomes. |
| doi_str_mv | 10.1073/pnas.0307550101 |
| format | Article |
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High tetA transcription caused a 10- to 80-fold decrease in the γδ resolution efficiency for the domain that includes the Tet module. Replacing tetA with genes encoding cytosolic proteins LacZ and Kan also caused the appearance of supercoil diffusion barriers in a defined region of the chromosome. In strains containing a functional TetR located next to a regulated lacZ reporter (PRtetR-PAlacZ), induction of transcription with chlortetracycline caused a 5-fold drop in resolution efficiency in the test domain interval. A short half-life resolves showed that barriers appeared and disappeared over a 10- to 20-min span. These studies demonstrate the importance of transcription in chromosome structure and the plasticity of supercoil domains in bacterial chromosomes.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.0307550101</identifier><identifier>PMID: 14993611</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Antiporters - genetics ; Antiporters - metabolism ; Bacteria ; Bacterial chromosomes ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Biochemistry ; Biological Sciences ; Chromosomes ; Chromosomes, Bacterial - chemistry ; Chromosomes, Bacterial - genetics ; Deoxyribonucleic acid ; DNA ; DNA, Bacterial - chemistry ; DNA, Bacterial - genetics ; DNA, Bacterial - metabolism ; DNA, Superhelical - chemistry ; DNA, Superhelical - genetics ; DNA, Superhelical - metabolism ; Genes ; Genes, Bacterial ; Membrane proteins ; Membranes ; Nucleic Acid Conformation ; Operons ; Phenotypes ; Plasmids ; Proteins ; Recombination, Genetic ; RNA ; Salmonella typhimurium ; Salmonella typhimurium - chemistry ; Salmonella typhimurium - genetics ; Salmonella typhimurium - metabolism ; TetA protein ; TetR protein ; Transcription, Genetic</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2004-03, Vol.101 (10), p.3398-3403</ispartof><rights>Copyright 1993/2004 The National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Mar 9, 2004</rights><rights>Copyright © 2004, The National Academy of Sciences 2004</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c524t-2113f130a2aa7c980c7c2eb39de5f01b788462cf0cddd2db2cbbe290c7f202a63</citedby><cites>FETCH-LOGICAL-c524t-2113f130a2aa7c980c7c2eb39de5f01b788462cf0cddd2db2cbbe290c7f202a63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/101/10.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/3371490$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/3371490$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,724,777,781,800,882,27905,27906,53772,53774,57998,58231</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14993611$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Deng, Shuang</creatorcontrib><creatorcontrib>Stein, Richard A.</creatorcontrib><creatorcontrib>Higgins, N. Patrick</creatorcontrib><creatorcontrib>Gottesman, Susan</creatorcontrib><title>Transcription-Induced Barriers to Supercoil Diffusion in the Salmonella typhimurium Chromosome</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Transcription and replication both influence and are influenced by superhelical changes in DNA. Explaining how supercoil movement is channeled in living chromosomes has been a major problem for 30 years. Transcription of membrane-associated proteins leads to localized hypersupercoiling of plasmid DNA, and this behavior indicates the presence of aberrant supercoil diffusion. Using the lambda Red recombination system, we constructed model domains in the Salmonella typhimurium chromosome to analyze supercoiling dynamics of regions encoding membrane proteins. Regulation of Tn10-derived tetracycline resistance involves a repressor, TetR, and a membrane-bound export pump, TetA. Strains deficient in TetR activity had 60-fold higher transcription levels (from PA) than TetR-positive strains. High tetA transcription caused a 10- to 80-fold decrease in the γδ resolution efficiency for the domain that includes the Tet module. Replacing tetA with genes encoding cytosolic proteins LacZ and Kan also caused the appearance of supercoil diffusion barriers in a defined region of the chromosome. In strains containing a functional TetR located next to a regulated lacZ reporter (PRtetR-PAlacZ), induction of transcription with chlortetracycline caused a 5-fold drop in resolution efficiency in the test domain interval. A short half-life resolves showed that barriers appeared and disappeared over a 10- to 20-min span. These studies demonstrate the importance of transcription in chromosome structure and the plasticity of supercoil domains in bacterial chromosomes.</description><subject>Antiporters - genetics</subject><subject>Antiporters - metabolism</subject><subject>Bacteria</subject><subject>Bacterial chromosomes</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Biochemistry</subject><subject>Biological Sciences</subject><subject>Chromosomes</subject><subject>Chromosomes, Bacterial - chemistry</subject><subject>Chromosomes, Bacterial - genetics</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Bacterial - chemistry</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA, Bacterial - metabolism</subject><subject>DNA, Superhelical - chemistry</subject><subject>DNA, Superhelical - genetics</subject><subject>DNA, Superhelical - metabolism</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>Membrane proteins</subject><subject>Membranes</subject><subject>Nucleic Acid Conformation</subject><subject>Operons</subject><subject>Phenotypes</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Recombination, Genetic</subject><subject>RNA</subject><subject>Salmonella typhimurium</subject><subject>Salmonella typhimurium - chemistry</subject><subject>Salmonella typhimurium - genetics</subject><subject>Salmonella typhimurium - metabolism</subject><subject>TetA protein</subject><subject>TetR protein</subject><subject>Transcription, Genetic</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1v1DAQxS1ERZfCmQuCiAPqJe3YTuL4wAEWKJUqcWi5YjmOw3qV2Kk_EP3vcbSrLnAoJx_8e2_ezEPoBYYzDIyez1aGM6DA6how4EdohYHjsqk4PEYrAMLKtiLVMXoawhYAeN3CE3SMK85pg_EKfb_x0gblzRyNs-Wl7ZPSffFBem-0D0V0xXWatVfOjMVHMwwpZK4wtogbXVzLcXJWj6Ms4t28MVPyJk3FeuPd5IKb9DN0NMgx6Of79wR9-_zpZv2lvPp6cbl-f1WqmlSxJBjTAVOQREqmeAuKKaI7yntdD4A71rZVQ9QAqu970ndEdZ0mPGMDASIbeoLe7Xzn1E26V9pGL0cxezNJfyecNOLvH2s24of7KSijFaNZ_3av9-426RDFZIJaFrPapSAYZgSaFv8XxDkqr1ibwTf_gFuXvM1HEARwBS2pl7HnO0h5F4LXw31iDGIpWCwFi0PBWfHqz0UP_L7RDLzeA4vyYIcXS0r5kuz0YUIMaRyj_hUz-nKHbkN0_p6llOV5QH8DNkTFiQ</recordid><startdate>20040309</startdate><enddate>20040309</enddate><creator>Deng, Shuang</creator><creator>Stein, Richard A.</creator><creator>Higgins, N. 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Patrick</au><au>Gottesman, Susan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Transcription-Induced Barriers to Supercoil Diffusion in the Salmonella typhimurium Chromosome</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>2004-03-09</date><risdate>2004</risdate><volume>101</volume><issue>10</issue><spage>3398</spage><epage>3403</epage><pages>3398-3403</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>Transcription and replication both influence and are influenced by superhelical changes in DNA. Explaining how supercoil movement is channeled in living chromosomes has been a major problem for 30 years. Transcription of membrane-associated proteins leads to localized hypersupercoiling of plasmid DNA, and this behavior indicates the presence of aberrant supercoil diffusion. Using the lambda Red recombination system, we constructed model domains in the Salmonella typhimurium chromosome to analyze supercoiling dynamics of regions encoding membrane proteins. Regulation of Tn10-derived tetracycline resistance involves a repressor, TetR, and a membrane-bound export pump, TetA. Strains deficient in TetR activity had 60-fold higher transcription levels (from PA) than TetR-positive strains. High tetA transcription caused a 10- to 80-fold decrease in the γδ resolution efficiency for the domain that includes the Tet module. Replacing tetA with genes encoding cytosolic proteins LacZ and Kan also caused the appearance of supercoil diffusion barriers in a defined region of the chromosome. In strains containing a functional TetR located next to a regulated lacZ reporter (PRtetR-PAlacZ), induction of transcription with chlortetracycline caused a 5-fold drop in resolution efficiency in the test domain interval. A short half-life resolves showed that barriers appeared and disappeared over a 10- to 20-min span. These studies demonstrate the importance of transcription in chromosome structure and the plasticity of supercoil domains in bacterial chromosomes.</abstract><cop>United States</cop><pub>National Academy of Sciences</pub><pmid>14993611</pmid><doi>10.1073/pnas.0307550101</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Antiporters - genetics Antiporters - metabolism Bacteria Bacterial chromosomes Bacterial Proteins - genetics Bacterial Proteins - metabolism Biochemistry Biological Sciences Chromosomes Chromosomes, Bacterial - chemistry Chromosomes, Bacterial - genetics Deoxyribonucleic acid DNA DNA, Bacterial - chemistry DNA, Bacterial - genetics DNA, Bacterial - metabolism DNA, Superhelical - chemistry DNA, Superhelical - genetics DNA, Superhelical - metabolism Genes Genes, Bacterial Membrane proteins Membranes Nucleic Acid Conformation Operons Phenotypes Plasmids Proteins Recombination, Genetic RNA Salmonella typhimurium Salmonella typhimurium - chemistry Salmonella typhimurium - genetics Salmonella typhimurium - metabolism TetA protein TetR protein Transcription, Genetic |
| title | Transcription-Induced Barriers to Supercoil Diffusion in the Salmonella typhimurium Chromosome |
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