The mdoA locus of Escherichia coli consists of an operon under osmotic control
Summary In Escherichia coli, the 5 kb mdoA locus is involved in the osmotically controlled biosynthesis of periplasmic membrane‐derived oligosaccharides (MDOs). The structure of this locus was analysed by in vitro cassette insertion, transposon mutagenesis, and gene‐fusion analysis. A ‘neo’ cassette...
Gespeichert in:
Veröffentlicht in: | Molecular microbiology 1991-07, Vol.5 (7), p.1745-1753 |
---|---|
Hauptverfasser: | , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 1753 |
---|---|
container_issue | 7 |
container_start_page | 1745 |
container_title | Molecular microbiology |
container_volume | 5 |
creator | Lacroix, J.‐M. Loubens, I. Tempête, M. Menichi, B. Bohin, J.‐P. |
description | Summary
In Escherichia coli, the 5 kb mdoA locus is involved in the osmotically controlled biosynthesis of periplasmic membrane‐derived oligosaccharides (MDOs). The structure of this locus was analysed by in vitro cassette insertion, transposon mutagenesis, and gene‐fusion analysis.
A ‘neo’ cassette, derived from the neomycin phosphotransferase II region of transposon Tn5, was inserted into mdoA, borne by a multicopy plasmid. This plasmid was shown to complement two previously described mdoA mutations, depending on the orientation of the exogenous gene. Thus, the gene altered by these mutations could be expressed under the control of the exogenous promoter. Moreover, the ‘neo’ cassette inactivated another, uncharacterlzed, mdo gene, because when this insertion was transferred into the chromosome MDO synthesis was abolished.
The existence of a second gene was confirmed by complementation analysis with a collection of Tn1000 insertions into mdoA. Two groups were defined, and the two genes are organized into an operon (mdoGH). This conclusion was reached because Tn1000 insertions in the first gene displayed a polar effect on the expression of the second gene.
An active gene fusion was obtained on a multicopy plasmid between the beginning of mdoH and lacZ. The hybrid β‐galactosidase activity followed the same osmotically controlled response as that described for of MDO synthesis. This regulation was unaffected by the presence, or absence, of MDOs In the periplasm. Finally, the amount of mdoA‐specific mRNAs, determined by dot blot hybridization, decreased when the osmolarity of the growth medium increased. |
doi_str_mv | 10.1111/j.1365-2958.1991.tb01924.x |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_16221474</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>16221474</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4965-212c131f557cb6e5484e279f2a628a9958527509fd456fb181de31f45438c4a33</originalsourceid><addsrcrecordid>eNqVkMtKxDAUhoMoOl4eQSgi7qbm5NI2bmSQ8QKjbhTchTRNmQxtMyYtOm9va4dxbRbJ4v9Ozs-H0AXgGPpzvYqBJnxKBM9iEALiNscgCIu_99BkF-2jCRYcT2lGPo7QcQgrjIHihB6iQ8goE0An6OVtaaK6cLOocroLkSujedBL461eWhVpV9n-aoIN7W-omsitjXdN1DWF8ZELtWutHpjWu-oUHZSqCuZs-56g9_v5293jdPH68HQ3W0w1E0M9IBoolJynOk8MZxkzJBUlUQnJlOjLc5JyLMqC8aTMIYPC9DjjjGaaKUpP0NX479q7z86EVtY2aFNVqjGuCxISQoClrAdvRlB7F4I3pVx7Wyu_kYDlIFOu5GBMDsbkIFNuZcrvfvh8u6XLa1P8jY72-vxym6ugVVV61WgbdhgTKRM86bHbEfuyldn8o4B8fn6ClHH6AzDGkCA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16221474</pqid></control><display><type>article</type><title>The mdoA locus of Escherichia coli consists of an operon under osmotic control</title><source>MEDLINE</source><source>Access via Wiley Online Library</source><creator>Lacroix, J.‐M. ; Loubens, I. ; Tempête, M. ; Menichi, B. ; Bohin, J.‐P.</creator><creatorcontrib>Lacroix, J.‐M. ; Loubens, I. ; Tempête, M. ; Menichi, B. ; Bohin, J.‐P.</creatorcontrib><description>Summary
In Escherichia coli, the 5 kb mdoA locus is involved in the osmotically controlled biosynthesis of periplasmic membrane‐derived oligosaccharides (MDOs). The structure of this locus was analysed by in vitro cassette insertion, transposon mutagenesis, and gene‐fusion analysis.
A ‘neo’ cassette, derived from the neomycin phosphotransferase II region of transposon Tn5, was inserted into mdoA, borne by a multicopy plasmid. This plasmid was shown to complement two previously described mdoA mutations, depending on the orientation of the exogenous gene. Thus, the gene altered by these mutations could be expressed under the control of the exogenous promoter. Moreover, the ‘neo’ cassette inactivated another, uncharacterlzed, mdo gene, because when this insertion was transferred into the chromosome MDO synthesis was abolished.
The existence of a second gene was confirmed by complementation analysis with a collection of Tn1000 insertions into mdoA. Two groups were defined, and the two genes are organized into an operon (mdoGH). This conclusion was reached because Tn1000 insertions in the first gene displayed a polar effect on the expression of the second gene.
An active gene fusion was obtained on a multicopy plasmid between the beginning of mdoH and lacZ. The hybrid β‐galactosidase activity followed the same osmotically controlled response as that described for of MDO synthesis. This regulation was unaffected by the presence, or absence, of MDOs In the periplasm. Finally, the amount of mdoA‐specific mRNAs, determined by dot blot hybridization, decreased when the osmolarity of the growth medium increased.</description><identifier>ISSN: 0950-382X</identifier><identifier>EISSN: 1365-2958</identifier><identifier>DOI: 10.1111/j.1365-2958.1991.tb01924.x</identifier><identifier>PMID: 1834913</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Bacteriology ; Biological and medical sciences ; DNA Mutational Analysis ; Escherichia coli ; Escherichia coli - genetics ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation, Bacterial ; Genetics ; Glucosyltransferases - genetics ; Microbiology ; Mutagenesis, Insertional ; Oligosaccharides - biosynthesis ; Oligosaccharides - genetics ; Oligosaccharides - isolation & purification ; Operon - genetics ; Osmotic Pressure ; Plasmids - genetics ; Recombinant Fusion Proteins ; Restriction Mapping ; RNA, Messenger - biosynthesis ; Sodium Chloride - pharmacology ; Transcription, Genetic</subject><ispartof>Molecular microbiology, 1991-07, Vol.5 (7), p.1745-1753</ispartof><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4965-212c131f557cb6e5484e279f2a628a9958527509fd456fb181de31f45438c4a33</citedby><cites>FETCH-LOGICAL-c4965-212c131f557cb6e5484e279f2a628a9958527509fd456fb181de31f45438c4a33</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.1365-2958.1991.tb01924.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1365-2958.1991.tb01924.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4974956$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1834913$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lacroix, J.‐M.</creatorcontrib><creatorcontrib>Loubens, I.</creatorcontrib><creatorcontrib>Tempête, M.</creatorcontrib><creatorcontrib>Menichi, B.</creatorcontrib><creatorcontrib>Bohin, J.‐P.</creatorcontrib><title>The mdoA locus of Escherichia coli consists of an operon under osmotic control</title><title>Molecular microbiology</title><addtitle>Mol Microbiol</addtitle><description>Summary
In Escherichia coli, the 5 kb mdoA locus is involved in the osmotically controlled biosynthesis of periplasmic membrane‐derived oligosaccharides (MDOs). The structure of this locus was analysed by in vitro cassette insertion, transposon mutagenesis, and gene‐fusion analysis.
A ‘neo’ cassette, derived from the neomycin phosphotransferase II region of transposon Tn5, was inserted into mdoA, borne by a multicopy plasmid. This plasmid was shown to complement two previously described mdoA mutations, depending on the orientation of the exogenous gene. Thus, the gene altered by these mutations could be expressed under the control of the exogenous promoter. Moreover, the ‘neo’ cassette inactivated another, uncharacterlzed, mdo gene, because when this insertion was transferred into the chromosome MDO synthesis was abolished.
The existence of a second gene was confirmed by complementation analysis with a collection of Tn1000 insertions into mdoA. Two groups were defined, and the two genes are organized into an operon (mdoGH). This conclusion was reached because Tn1000 insertions in the first gene displayed a polar effect on the expression of the second gene.
An active gene fusion was obtained on a multicopy plasmid between the beginning of mdoH and lacZ. The hybrid β‐galactosidase activity followed the same osmotically controlled response as that described for of MDO synthesis. This regulation was unaffected by the presence, or absence, of MDOs In the periplasm. Finally, the amount of mdoA‐specific mRNAs, determined by dot blot hybridization, decreased when the osmolarity of the growth medium increased.</description><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>DNA Mutational Analysis</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Genetics</subject><subject>Glucosyltransferases - genetics</subject><subject>Microbiology</subject><subject>Mutagenesis, Insertional</subject><subject>Oligosaccharides - biosynthesis</subject><subject>Oligosaccharides - genetics</subject><subject>Oligosaccharides - isolation & purification</subject><subject>Operon - genetics</subject><subject>Osmotic Pressure</subject><subject>Plasmids - genetics</subject><subject>Recombinant Fusion Proteins</subject><subject>Restriction Mapping</subject><subject>RNA, Messenger - biosynthesis</subject><subject>Sodium Chloride - pharmacology</subject><subject>Transcription, Genetic</subject><issn>0950-382X</issn><issn>1365-2958</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkMtKxDAUhoMoOl4eQSgi7qbm5NI2bmSQ8QKjbhTchTRNmQxtMyYtOm9va4dxbRbJ4v9Ozs-H0AXgGPpzvYqBJnxKBM9iEALiNscgCIu_99BkF-2jCRYcT2lGPo7QcQgrjIHihB6iQ8goE0An6OVtaaK6cLOocroLkSujedBL461eWhVpV9n-aoIN7W-omsitjXdN1DWF8ZELtWutHpjWu-oUHZSqCuZs-56g9_v5293jdPH68HQ3W0w1E0M9IBoolJynOk8MZxkzJBUlUQnJlOjLc5JyLMqC8aTMIYPC9DjjjGaaKUpP0NX479q7z86EVtY2aFNVqjGuCxISQoClrAdvRlB7F4I3pVx7Wyu_kYDlIFOu5GBMDsbkIFNuZcrvfvh8u6XLa1P8jY72-vxym6ugVVV61WgbdhgTKRM86bHbEfuyldn8o4B8fn6ClHH6AzDGkCA</recordid><startdate>199107</startdate><enddate>199107</enddate><creator>Lacroix, J.‐M.</creator><creator>Loubens, I.</creator><creator>Tempête, M.</creator><creator>Menichi, B.</creator><creator>Bohin, J.‐P.</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>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>199107</creationdate><title>The mdoA locus of Escherichia coli consists of an operon under osmotic control</title><author>Lacroix, J.‐M. ; Loubens, I. ; Tempête, M. ; Menichi, B. ; Bohin, J.‐P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4965-212c131f557cb6e5484e279f2a628a9958527509fd456fb181de31f45438c4a33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>DNA Mutational Analysis</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation, Bacterial</topic><topic>Genetics</topic><topic>Glucosyltransferases - genetics</topic><topic>Microbiology</topic><topic>Mutagenesis, Insertional</topic><topic>Oligosaccharides - biosynthesis</topic><topic>Oligosaccharides - genetics</topic><topic>Oligosaccharides - isolation & purification</topic><topic>Operon - genetics</topic><topic>Osmotic Pressure</topic><topic>Plasmids - genetics</topic><topic>Recombinant Fusion Proteins</topic><topic>Restriction Mapping</topic><topic>RNA, Messenger - biosynthesis</topic><topic>Sodium Chloride - pharmacology</topic><topic>Transcription, Genetic</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lacroix, J.‐M.</creatorcontrib><creatorcontrib>Loubens, I.</creatorcontrib><creatorcontrib>Tempête, M.</creatorcontrib><creatorcontrib>Menichi, B.</creatorcontrib><creatorcontrib>Bohin, J.‐P.</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>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><jtitle>Molecular microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lacroix, J.‐M.</au><au>Loubens, I.</au><au>Tempête, M.</au><au>Menichi, B.</au><au>Bohin, J.‐P.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The mdoA locus of Escherichia coli consists of an operon under osmotic control</atitle><jtitle>Molecular microbiology</jtitle><addtitle>Mol Microbiol</addtitle><date>1991-07</date><risdate>1991</risdate><volume>5</volume><issue>7</issue><spage>1745</spage><epage>1753</epage><pages>1745-1753</pages><issn>0950-382X</issn><eissn>1365-2958</eissn><abstract>Summary
In Escherichia coli, the 5 kb mdoA locus is involved in the osmotically controlled biosynthesis of periplasmic membrane‐derived oligosaccharides (MDOs). The structure of this locus was analysed by in vitro cassette insertion, transposon mutagenesis, and gene‐fusion analysis.
A ‘neo’ cassette, derived from the neomycin phosphotransferase II region of transposon Tn5, was inserted into mdoA, borne by a multicopy plasmid. This plasmid was shown to complement two previously described mdoA mutations, depending on the orientation of the exogenous gene. Thus, the gene altered by these mutations could be expressed under the control of the exogenous promoter. Moreover, the ‘neo’ cassette inactivated another, uncharacterlzed, mdo gene, because when this insertion was transferred into the chromosome MDO synthesis was abolished.
The existence of a second gene was confirmed by complementation analysis with a collection of Tn1000 insertions into mdoA. Two groups were defined, and the two genes are organized into an operon (mdoGH). This conclusion was reached because Tn1000 insertions in the first gene displayed a polar effect on the expression of the second gene.
An active gene fusion was obtained on a multicopy plasmid between the beginning of mdoH and lacZ. The hybrid β‐galactosidase activity followed the same osmotically controlled response as that described for of MDO synthesis. This regulation was unaffected by the presence, or absence, of MDOs In the periplasm. Finally, the amount of mdoA‐specific mRNAs, determined by dot blot hybridization, decreased when the osmolarity of the growth medium increased.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>1834913</pmid><doi>10.1111/j.1365-2958.1991.tb01924.x</doi><tpages>9</tpages></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0950-382X |
ispartof | Molecular microbiology, 1991-07, Vol.5 (7), p.1745-1753 |
issn | 0950-382X 1365-2958 |
language | eng |
recordid | cdi_proquest_miscellaneous_16221474 |
source | MEDLINE; Access via Wiley Online Library |
subjects | Bacteriology Biological and medical sciences DNA Mutational Analysis Escherichia coli Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Bacterial Genetics Glucosyltransferases - genetics Microbiology Mutagenesis, Insertional Oligosaccharides - biosynthesis Oligosaccharides - genetics Oligosaccharides - isolation & purification Operon - genetics Osmotic Pressure Plasmids - genetics Recombinant Fusion Proteins Restriction Mapping RNA, Messenger - biosynthesis Sodium Chloride - pharmacology Transcription, Genetic |
title | The mdoA locus of Escherichia coli consists of an operon under osmotic control |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T10%3A28%3A01IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=The%20mdoA%20locus%20of%20Escherichia%20coli%20consists%20of%20an%20operon%20under%20osmotic%20control&rft.jtitle=Molecular%20microbiology&rft.au=Lacroix,%20J.%E2%80%90M.&rft.date=1991-07&rft.volume=5&rft.issue=7&rft.spage=1745&rft.epage=1753&rft.pages=1745-1753&rft.issn=0950-382X&rft.eissn=1365-2958&rft_id=info:doi/10.1111/j.1365-2958.1991.tb01924.x&rft_dat=%3Cproquest_cross%3E16221474%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=16221474&rft_id=info:pmid/1834913&rfr_iscdi=true |