Transposon-mediated insertional mutagenesis of the D-alanyl-lipoteichoic acid ( dlt) operon raises methicillin resistance in Staphylococcus aureus
Two independent mutants of methicillin-resistant Staphylococcus aureus (MRSA), KAN96H1 and KAN96H2, were isolated by insertional mutagenesis with conjugative transposon Tn 918. In both, the minimal inhibitory concentration (MIC) of methicillin was increased to 128 compared to 16 mg/L for the parenta...
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| creator | Nakao, Ayumi Imai, Shin-ichiro Takano, Toshiya |
| description | Two independent mutants of methicillin-resistant
Staphylococcus aureus (MRSA), KAN96H1 and KAN96H2, were isolated by insertional mutagenesis with conjugative transposon Tn
918. In both, the minimal inhibitory concentration (MIC) of methicillin was increased to 128 compared to 16 mg/L for the parental strain KAN96. By transduction experiments, we verified that the insertion of Tn
918 conferred higher methicillin resistance on KAN96H1, but not on KAN96H2. In KAN96H1, the integration site of Tn
918 was located in the 6.1-kb
HindIII fragment of the KAN96 chromosomal DNA. We identified a novel D-alanyl-lipoteichoic acid (
dlt) operon of
S. aureus in this fragment. The amino acid sequences of four open reading frames of this operon were highly homologous to those of the
dlt operon genes of
Bacillus subtilis. The nucleotide sequence of the staphylococcal
dlt operon is under the accession number D86240 in the DDBJ/GenBank/EMBL databases. In KAN96H1, Tn
918 was inserted in the 5'-terminal region of the putative
dltB gene which encoded a hypothetical membrane transporter.
dlt transcripts of 4.7 kb were detected in KAN96, but were truncated to 2.3 kb in KAN96H1. No corresponding transcripts were observed in KAN96H2. Our results clearly demonstrated that defects in functions of the putative
dlt operon resulted in increased methicillin resistance in MRSA. |
| doi_str_mv | 10.1016/S0923-2508(00)01148-7 |
| format | Article |
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Staphylococcus aureus (MRSA), KAN96H1 and KAN96H2, were isolated by insertional mutagenesis with conjugative transposon Tn
918. In both, the minimal inhibitory concentration (MIC) of methicillin was increased to 128 compared to 16 mg/L for the parental strain KAN96. By transduction experiments, we verified that the insertion of Tn
918 conferred higher methicillin resistance on KAN96H1, but not on KAN96H2. In KAN96H1, the integration site of Tn
918 was located in the 6.1-kb
HindIII fragment of the KAN96 chromosomal DNA. We identified a novel D-alanyl-lipoteichoic acid (
dlt) operon of
S. aureus in this fragment. The amino acid sequences of four open reading frames of this operon were highly homologous to those of the
dlt operon genes of
Bacillus subtilis. The nucleotide sequence of the staphylococcal
dlt operon is under the accession number D86240 in the DDBJ/GenBank/EMBL databases. In KAN96H1, Tn
918 was inserted in the 5'-terminal region of the putative
dltB gene which encoded a hypothetical membrane transporter.
dlt transcripts of 4.7 kb were detected in KAN96, but were truncated to 2.3 kb in KAN96H1. No corresponding transcripts were observed in KAN96H2. Our results clearly demonstrated that defects in functions of the putative
dlt operon resulted in increased methicillin resistance in MRSA.</description><identifier>ISSN: 0923-2508</identifier><identifier>EISSN: 1769-7123</identifier><identifier>DOI: 10.1016/S0923-2508(00)01148-7</identifier><identifier>PMID: 11191807</identifier><language>eng</language><publisher>Paris: Elsevier SAS</publisher><subject>Amino Acid Sequence ; Antibacterial agents ; Antibiotics. Antiinfectious agents. Antiparasitic agents ; Bacteriology ; Biological and medical sciences ; D-Alanyl-lipoteichoic acid ; dlt gene ; dlt operon ; DNA Transposable Elements ; Drug Resistance, Microbial ; Fundamental and applied biological sciences. Psychology ; Genetics ; Humans ; insertional mutagenesis ; Medical sciences ; Methicillin - pharmacology ; methicillin resistance ; Microbial Sensitivity Tests ; Microbiology ; Molecular Sequence Data ; Mutagenesis, Insertional ; Operon ; Penicillins - pharmacology ; Pharmacology. Drug treatments ; Sequence Homology, Amino Acid ; Staphylococcus aureus ; Staphylococcus aureus - chemistry ; Staphylococcus aureus - drug effects ; Staphylococcus aureus - genetics ; Teichoic Acids - genetics ; transposon Tn918</subject><ispartof>Research in microbiology, 2000-12, Vol.151 (10), p.823-829</ispartof><rights>2000 Éditions scientifiques et médicales Elsevier SAS</rights><rights>2001 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0923-2508(00)01148-7$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=871207$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11191807$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nakao, Ayumi</creatorcontrib><creatorcontrib>Imai, Shin-ichiro</creatorcontrib><creatorcontrib>Takano, Toshiya</creatorcontrib><title>Transposon-mediated insertional mutagenesis of the D-alanyl-lipoteichoic acid ( dlt) operon raises methicillin resistance in Staphylococcus aureus</title><title>Research in microbiology</title><addtitle>Res Microbiol</addtitle><description>Two independent mutants of methicillin-resistant
Staphylococcus aureus (MRSA), KAN96H1 and KAN96H2, were isolated by insertional mutagenesis with conjugative transposon Tn
918. In both, the minimal inhibitory concentration (MIC) of methicillin was increased to 128 compared to 16 mg/L for the parental strain KAN96. By transduction experiments, we verified that the insertion of Tn
918 conferred higher methicillin resistance on KAN96H1, but not on KAN96H2. In KAN96H1, the integration site of Tn
918 was located in the 6.1-kb
HindIII fragment of the KAN96 chromosomal DNA. We identified a novel D-alanyl-lipoteichoic acid (
dlt) operon of
S. aureus in this fragment. The amino acid sequences of four open reading frames of this operon were highly homologous to those of the
dlt operon genes of
Bacillus subtilis. The nucleotide sequence of the staphylococcal
dlt operon is under the accession number D86240 in the DDBJ/GenBank/EMBL databases. In KAN96H1, Tn
918 was inserted in the 5'-terminal region of the putative
dltB gene which encoded a hypothetical membrane transporter.
dlt transcripts of 4.7 kb were detected in KAN96, but were truncated to 2.3 kb in KAN96H1. No corresponding transcripts were observed in KAN96H2. Our results clearly demonstrated that defects in functions of the putative
dlt operon resulted in increased methicillin resistance in MRSA.</description><subject>Amino Acid Sequence</subject><subject>Antibacterial agents</subject><subject>Antibiotics. Antiinfectious agents. Antiparasitic agents</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>D-Alanyl-lipoteichoic acid</subject><subject>dlt gene</subject><subject>dlt operon</subject><subject>DNA Transposable Elements</subject><subject>Drug Resistance, Microbial</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetics</subject><subject>Humans</subject><subject>insertional mutagenesis</subject><subject>Medical sciences</subject><subject>Methicillin - pharmacology</subject><subject>methicillin resistance</subject><subject>Microbial Sensitivity Tests</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Insertional</subject><subject>Operon</subject><subject>Penicillins - pharmacology</subject><subject>Pharmacology. Drug treatments</subject><subject>Sequence Homology, Amino Acid</subject><subject>Staphylococcus aureus</subject><subject>Staphylococcus aureus - chemistry</subject><subject>Staphylococcus aureus - drug effects</subject><subject>Staphylococcus aureus - genetics</subject><subject>Teichoic Acids - genetics</subject><subject>transposon Tn918</subject><issn>0923-2508</issn><issn>1769-7123</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kduKFDEQhoMo7uzqIygBQXYvWnPodNJXIut6gAUvdr0OmXS1U5LutElamNfwic3MjntVUPVRRf0fIa84e8cZ797fsV7IRihmLhm7Ypy3ptFPyIbrrm80F_Ip2TwiZ-Q851-McaV1-5yccc57bpjekL_3yc15iTnOzQQDugIDxTlDKhhnF-i0FvcTZsiYaRxp2QH91Ljg5n1oAi6xAPpdRE-dx4Fe0iGUKxoXSHGmyWGGTCcoO_QYAtbWYVFxs4d6hd4Vt-z2Ifro_ZqpWxOs-QV5NrqQ4eWpXpAfn2_ur782t9-_fLv-eNuAkLo0vQCpHB-kAqWVNqNzWwleSNBdZ5Tqfd-2YtiaOpZS-3Hsdcsk32rR98p08oK8fdi7pPh7hVzshNlDqL9BXLPl2rSyFaqCr0_guq0Z2SXh5NLe_k-xAm9OgMvehbFG6jE_cqbqOFIfHiioT_1BSDZ7hJrEgAl8sUNEy5k92LVHu_agzjJmj3atlv8AdmiYPQ</recordid><startdate>200012</startdate><enddate>200012</enddate><creator>Nakao, Ayumi</creator><creator>Imai, Shin-ichiro</creator><creator>Takano, Toshiya</creator><general>Elsevier SAS</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7QL</scope><scope>C1K</scope></search><sort><creationdate>200012</creationdate><title>Transposon-mediated insertional mutagenesis of the D-alanyl-lipoteichoic acid ( dlt) operon raises methicillin resistance in Staphylococcus aureus</title><author>Nakao, Ayumi ; Imai, Shin-ichiro ; Takano, Toshiya</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-e237t-92e35a1d35e57578faab3ec23e7668559c9442db8e57337cff974031b72995863</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Amino Acid Sequence</topic><topic>Antibacterial agents</topic><topic>Antibiotics. Antiinfectious agents. Antiparasitic agents</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>D-Alanyl-lipoteichoic acid</topic><topic>dlt gene</topic><topic>dlt operon</topic><topic>DNA Transposable Elements</topic><topic>Drug Resistance, Microbial</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genetics</topic><topic>Humans</topic><topic>insertional mutagenesis</topic><topic>Medical sciences</topic><topic>Methicillin - pharmacology</topic><topic>methicillin resistance</topic><topic>Microbial Sensitivity Tests</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis, Insertional</topic><topic>Operon</topic><topic>Penicillins - pharmacology</topic><topic>Pharmacology. Drug treatments</topic><topic>Sequence Homology, Amino Acid</topic><topic>Staphylococcus aureus</topic><topic>Staphylococcus aureus - chemistry</topic><topic>Staphylococcus aureus - drug effects</topic><topic>Staphylococcus aureus - genetics</topic><topic>Teichoic Acids - genetics</topic><topic>transposon Tn918</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nakao, Ayumi</creatorcontrib><creatorcontrib>Imai, Shin-ichiro</creatorcontrib><creatorcontrib>Takano, Toshiya</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Research in microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nakao, Ayumi</au><au>Imai, Shin-ichiro</au><au>Takano, Toshiya</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Transposon-mediated insertional mutagenesis of the D-alanyl-lipoteichoic acid ( dlt) operon raises methicillin resistance in Staphylococcus aureus</atitle><jtitle>Research in microbiology</jtitle><addtitle>Res Microbiol</addtitle><date>2000-12</date><risdate>2000</risdate><volume>151</volume><issue>10</issue><spage>823</spage><epage>829</epage><pages>823-829</pages><issn>0923-2508</issn><eissn>1769-7123</eissn><abstract>Two independent mutants of methicillin-resistant
Staphylococcus aureus (MRSA), KAN96H1 and KAN96H2, were isolated by insertional mutagenesis with conjugative transposon Tn
918. In both, the minimal inhibitory concentration (MIC) of methicillin was increased to 128 compared to 16 mg/L for the parental strain KAN96. By transduction experiments, we verified that the insertion of Tn
918 conferred higher methicillin resistance on KAN96H1, but not on KAN96H2. In KAN96H1, the integration site of Tn
918 was located in the 6.1-kb
HindIII fragment of the KAN96 chromosomal DNA. We identified a novel D-alanyl-lipoteichoic acid (
dlt) operon of
S. aureus in this fragment. The amino acid sequences of four open reading frames of this operon were highly homologous to those of the
dlt operon genes of
Bacillus subtilis. The nucleotide sequence of the staphylococcal
dlt operon is under the accession number D86240 in the DDBJ/GenBank/EMBL databases. In KAN96H1, Tn
918 was inserted in the 5'-terminal region of the putative
dltB gene which encoded a hypothetical membrane transporter.
dlt transcripts of 4.7 kb were detected in KAN96, but were truncated to 2.3 kb in KAN96H1. No corresponding transcripts were observed in KAN96H2. Our results clearly demonstrated that defects in functions of the putative
dlt operon resulted in increased methicillin resistance in MRSA.</abstract><cop>Paris</cop><pub>Elsevier SAS</pub><pmid>11191807</pmid><doi>10.1016/S0923-2508(00)01148-7</doi><tpages>7</tpages></addata></record> |
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| issn | 0923-2508 1769-7123 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Amino Acid Sequence Antibacterial agents Antibiotics. Antiinfectious agents. Antiparasitic agents Bacteriology Biological and medical sciences D-Alanyl-lipoteichoic acid dlt gene dlt operon DNA Transposable Elements Drug Resistance, Microbial Fundamental and applied biological sciences. Psychology Genetics Humans insertional mutagenesis Medical sciences Methicillin - pharmacology methicillin resistance Microbial Sensitivity Tests Microbiology Molecular Sequence Data Mutagenesis, Insertional Operon Penicillins - pharmacology Pharmacology. Drug treatments Sequence Homology, Amino Acid Staphylococcus aureus Staphylococcus aureus - chemistry Staphylococcus aureus - drug effects Staphylococcus aureus - genetics Teichoic Acids - genetics transposon Tn918 |
| title | Transposon-mediated insertional mutagenesis of the D-alanyl-lipoteichoic acid ( dlt) operon raises methicillin resistance in Staphylococcus aureus |
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