Fatty acid kinase A is an important determinant of biofilm formation in Staphylococcus aureus USA300
Methicillin-resistant Staphylococcus aureus (MRSA)-USA300 is notorious for its ability to cause community- and healthcare-acquired infections, which are even more difficult to treat when associated with a biofilm phenotype. We aimed to characterize the genetic determinants of biofilm formation in a...
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| Veröffentlicht in: | BMC genomics 2015-10, Vol.16 (1), p.861-861, Article 861 |
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| creator | Sabirova, J S Hernalsteens, J-P De Backer, S Xavier, B B Moons, P Turlej-Rogacka, A De Greve, H Goossens, H Malhotra-Kumar, S |
| description | Methicillin-resistant Staphylococcus aureus (MRSA)-USA300 is notorious for its ability to cause community- and healthcare-acquired infections, which are even more difficult to treat when associated with a biofilm phenotype. We aimed to characterize the genetic determinants of biofilm formation in a USA300 skin abscess isolate (UAS391) that formed prolific biofilms.
USA300 S. aureus strains, TCH1516 and FPR3757, were found to be closely related based on whole genome mapping (Argus™ Optical Mapping System, Opgen Inc, Gaithersburg, USA) to UAS391 (96.3-99.1 % similarity, P=0.0151), however differed markedly in biofilm formation (P=0.0001) on a dynamic assay (BioFlux 200, Fluxion Biosciences, USA). Comparison of whole genome sequences of these strains identified differences in a total of 18 genes. Corresponding Tn (bursa aurealis-bearing) knockout mutants in these target genes were obtained from a publicly available mutant library of the same clonal lineage (USA300-JE2) and were characterized phenotypically for biofilm formation. Tn mutants showing significant differences in biofilm formation were utilized for transduction into a plasmid-cured erythromycin-sensitive derivative of UAS391 and for complementation experiments. All strains were tested on the dynamic assay, and 17h-biofilms were stained (SYTO9, Life Technologies) and fluorescence intensity quantified by microscopy (Zeiss, ImageJ). Gene expression levels in Tn and transduced mutants were studied by quantitative reverse transcriptase PCR (StepOnePlusTM, Applied Biosystems®).
Comparison of the sequenced genomes of TCH1516, FPR3757 and UAS391 yielded a limited number of variant genes (n=18) that were hypothesized to account for the observed difference in biofilm-forming capacity. Screening of Tn mutants disrupted in these target genes identified one mutant (NE229) bearing a transposon insertion in SAUSA300_1119 (fakA), which exhibited increased biofilm formation similar to UAS391 (P=0.9320). Transduction experiments confirmed that fakA::Tn corresponded to 1.9- to 4.6-fold increase in biofilm formation depending on the USA300 strain background (P≤0.0007), while complementation of the TCH1516 wild-type fakA allele in UAS391 resulted in a 4.3-fold reduction in biofilm formation (P |
| doi_str_mv | 10.1186/s12864-015-1956-8 |
| format | Article |
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USA300 S. aureus strains, TCH1516 and FPR3757, were found to be closely related based on whole genome mapping (Argus™ Optical Mapping System, Opgen Inc, Gaithersburg, USA) to UAS391 (96.3-99.1 % similarity, P=0.0151), however differed markedly in biofilm formation (P=0.0001) on a dynamic assay (BioFlux 200, Fluxion Biosciences, USA). Comparison of whole genome sequences of these strains identified differences in a total of 18 genes. Corresponding Tn (bursa aurealis-bearing) knockout mutants in these target genes were obtained from a publicly available mutant library of the same clonal lineage (USA300-JE2) and were characterized phenotypically for biofilm formation. Tn mutants showing significant differences in biofilm formation were utilized for transduction into a plasmid-cured erythromycin-sensitive derivative of UAS391 and for complementation experiments. All strains were tested on the dynamic assay, and 17h-biofilms were stained (SYTO9, Life Technologies) and fluorescence intensity quantified by microscopy (Zeiss, ImageJ). Gene expression levels in Tn and transduced mutants were studied by quantitative reverse transcriptase PCR (StepOnePlusTM, Applied Biosystems®).
Comparison of the sequenced genomes of TCH1516, FPR3757 and UAS391 yielded a limited number of variant genes (n=18) that were hypothesized to account for the observed difference in biofilm-forming capacity. Screening of Tn mutants disrupted in these target genes identified one mutant (NE229) bearing a transposon insertion in SAUSA300_1119 (fakA), which exhibited increased biofilm formation similar to UAS391 (P=0.9320). Transduction experiments confirmed that fakA::Tn corresponded to 1.9- to 4.6-fold increase in biofilm formation depending on the USA300 strain background (P≤0.0007), while complementation of the TCH1516 wild-type fakA allele in UAS391 resulted in a 4.3-fold reduction in biofilm formation (P<0.0001).
This sequential approach, consisting of strain typing, genome comparison and functional genomics, identified fakA, a recently described fatty acid kinase in S. aureus that is essential for phospholipid synthesis and also impacts the transcription of numerous virulence factors, as a negative regulator of biofilm formation in S. aureus USA300.</description><identifier>ISSN: 1471-2164</identifier><identifier>EISSN: 1471-2164</identifier><identifier>DOI: 10.1186/s12864-015-1956-8</identifier><identifier>PMID: 26502874</identifier><language>eng</language><publisher>England: BioMed Central Ltd</publisher><subject>Abscess ; Anopheles ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Biofilms - growth & development ; Comparative analysis ; Genetic aspects ; Genome, Bacterial - genetics ; Genomics ; Growth ; Health aspects ; Mutation ; Plasmids - genetics ; Staphylococcus aureus - enzymology ; Staphylococcus aureus - genetics ; Staphylococcus aureus - physiology ; Transposons</subject><ispartof>BMC genomics, 2015-10, Vol.16 (1), p.861-861, Article 861</ispartof><rights>COPYRIGHT 2015 BioMed Central Ltd.</rights><rights>Copyright BioMed Central 2015</rights><rights>Sabirova et al. 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c598t-71088fe15858207dcffcf810da679731c513f9eb5c5d03279ed36ea1bc114e8a3</citedby><cites>FETCH-LOGICAL-c598t-71088fe15858207dcffcf810da679731c513f9eb5c5d03279ed36ea1bc114e8a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4623894/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4623894/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26502874$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sabirova, J S</creatorcontrib><creatorcontrib>Hernalsteens, J-P</creatorcontrib><creatorcontrib>De Backer, S</creatorcontrib><creatorcontrib>Xavier, B B</creatorcontrib><creatorcontrib>Moons, P</creatorcontrib><creatorcontrib>Turlej-Rogacka, A</creatorcontrib><creatorcontrib>De Greve, H</creatorcontrib><creatorcontrib>Goossens, H</creatorcontrib><creatorcontrib>Malhotra-Kumar, S</creatorcontrib><title>Fatty acid kinase A is an important determinant of biofilm formation in Staphylococcus aureus USA300</title><title>BMC genomics</title><addtitle>BMC Genomics</addtitle><description>Methicillin-resistant Staphylococcus aureus (MRSA)-USA300 is notorious for its ability to cause community- and healthcare-acquired infections, which are even more difficult to treat when associated with a biofilm phenotype. We aimed to characterize the genetic determinants of biofilm formation in a USA300 skin abscess isolate (UAS391) that formed prolific biofilms.
USA300 S. aureus strains, TCH1516 and FPR3757, were found to be closely related based on whole genome mapping (Argus™ Optical Mapping System, Opgen Inc, Gaithersburg, USA) to UAS391 (96.3-99.1 % similarity, P=0.0151), however differed markedly in biofilm formation (P=0.0001) on a dynamic assay (BioFlux 200, Fluxion Biosciences, USA). Comparison of whole genome sequences of these strains identified differences in a total of 18 genes. Corresponding Tn (bursa aurealis-bearing) knockout mutants in these target genes were obtained from a publicly available mutant library of the same clonal lineage (USA300-JE2) and were characterized phenotypically for biofilm formation. Tn mutants showing significant differences in biofilm formation were utilized for transduction into a plasmid-cured erythromycin-sensitive derivative of UAS391 and for complementation experiments. All strains were tested on the dynamic assay, and 17h-biofilms were stained (SYTO9, Life Technologies) and fluorescence intensity quantified by microscopy (Zeiss, ImageJ). Gene expression levels in Tn and transduced mutants were studied by quantitative reverse transcriptase PCR (StepOnePlusTM, Applied Biosystems®).
Comparison of the sequenced genomes of TCH1516, FPR3757 and UAS391 yielded a limited number of variant genes (n=18) that were hypothesized to account for the observed difference in biofilm-forming capacity. Screening of Tn mutants disrupted in these target genes identified one mutant (NE229) bearing a transposon insertion in SAUSA300_1119 (fakA), which exhibited increased biofilm formation similar to UAS391 (P=0.9320). Transduction experiments confirmed that fakA::Tn corresponded to 1.9- to 4.6-fold increase in biofilm formation depending on the USA300 strain background (P≤0.0007), while complementation of the TCH1516 wild-type fakA allele in UAS391 resulted in a 4.3-fold reduction in biofilm formation (P<0.0001).
This sequential approach, consisting of strain typing, genome comparison and functional genomics, identified fakA, a recently described fatty acid kinase in S. aureus that is essential for phospholipid synthesis and also impacts the transcription of numerous virulence factors, as a negative regulator of biofilm formation in S. aureus USA300.</description><subject>Abscess</subject><subject>Anopheles</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Biofilms - growth & development</subject><subject>Comparative analysis</subject><subject>Genetic aspects</subject><subject>Genome, Bacterial - genetics</subject><subject>Genomics</subject><subject>Growth</subject><subject>Health aspects</subject><subject>Mutation</subject><subject>Plasmids - genetics</subject><subject>Staphylococcus aureus - enzymology</subject><subject>Staphylococcus aureus - genetics</subject><subject>Staphylococcus aureus - physiology</subject><subject>Transposons</subject><issn>1471-2164</issn><issn>1471-2164</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNptkl9rFDEUxQdR7B_9AL5IwBf7MDV3MplkXoSlWC0UBNc-h2zmZps6k6xJprjf3ixba1ckDzdwf-dccnOq6g3QcwDZfUjQyK6tKfAaet7V8ll1DK2AuoGuff7kflSdpHRHKQjZ8JfVUdNx2kjRHlfDpc55S7RxA_nhvE5IFsQloj1x0ybErH0mA2aMU-mWe7Bk5YJ140RsiJPOLhTUk2XWm9vtGEwwZi76OWIpN8sFo_RV9cLqMeHrh3pa3Vx--n7xpb7--vnqYnFdG97LXAugUloELrlsqBiMtcZKoIPuRC8YGA7M9rjihg-UNaLHgXWoYWUAWpSanVYf976beTXhYNDnqEe1iW7ScauCduqw492tWod71XYNk31bDN4_GMTwc8aU1eSSwXHUHsOcFIgytW-khIK--we9C3P05XmFEr1kvHzNX2qtR1TO21Dmmp2pWvAWWNeC6At1_h-qnAEnZ4LHsm48FJwdCAqT8Vde6zkldbX8dsjCnjUxpBTRPu4DqNrFSO1jpEqM1C5GShbN26eLfFT8yQ37DVm1wRE</recordid><startdate>20151026</startdate><enddate>20151026</enddate><creator>Sabirova, J S</creator><creator>Hernalsteens, J-P</creator><creator>De Backer, S</creator><creator>Xavier, B B</creator><creator>Moons, P</creator><creator>Turlej-Rogacka, A</creator><creator>De Greve, H</creator><creator>Goossens, H</creator><creator>Malhotra-Kumar, S</creator><general>BioMed Central Ltd</general><general>BioMed Central</general><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>ISR</scope><scope>3V.</scope><scope>7QP</scope><scope>7QR</scope><scope>7SS</scope><scope>7TK</scope><scope>7U7</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20151026</creationdate><title>Fatty acid kinase A is an important determinant of biofilm formation in Staphylococcus aureus USA300</title><author>Sabirova, J S ; Hernalsteens, J-P ; De Backer, S ; Xavier, B B ; Moons, P ; Turlej-Rogacka, A ; De Greve, H ; Goossens, H ; Malhotra-Kumar, S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c598t-71088fe15858207dcffcf810da679731c513f9eb5c5d03279ed36ea1bc114e8a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Abscess</topic><topic>Anopheles</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Biofilms - growth & development</topic><topic>Comparative analysis</topic><topic>Genetic aspects</topic><topic>Genome, Bacterial - genetics</topic><topic>Genomics</topic><topic>Growth</topic><topic>Health aspects</topic><topic>Mutation</topic><topic>Plasmids - genetics</topic><topic>Staphylococcus aureus - enzymology</topic><topic>Staphylococcus aureus - genetics</topic><topic>Staphylococcus aureus - physiology</topic><topic>Transposons</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sabirova, J S</creatorcontrib><creatorcontrib>Hernalsteens, J-P</creatorcontrib><creatorcontrib>De Backer, S</creatorcontrib><creatorcontrib>Xavier, B B</creatorcontrib><creatorcontrib>Moons, P</creatorcontrib><creatorcontrib>Turlej-Rogacka, A</creatorcontrib><creatorcontrib>De Greve, H</creatorcontrib><creatorcontrib>Goossens, H</creatorcontrib><creatorcontrib>Malhotra-Kumar, S</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Neurosciences Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>BMC genomics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sabirova, J S</au><au>Hernalsteens, J-P</au><au>De Backer, S</au><au>Xavier, B B</au><au>Moons, P</au><au>Turlej-Rogacka, A</au><au>De Greve, H</au><au>Goossens, H</au><au>Malhotra-Kumar, S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Fatty acid kinase A is an important determinant of biofilm formation in Staphylococcus aureus USA300</atitle><jtitle>BMC genomics</jtitle><addtitle>BMC Genomics</addtitle><date>2015-10-26</date><risdate>2015</risdate><volume>16</volume><issue>1</issue><spage>861</spage><epage>861</epage><pages>861-861</pages><artnum>861</artnum><issn>1471-2164</issn><eissn>1471-2164</eissn><abstract>Methicillin-resistant Staphylococcus aureus (MRSA)-USA300 is notorious for its ability to cause community- and healthcare-acquired infections, which are even more difficult to treat when associated with a biofilm phenotype. We aimed to characterize the genetic determinants of biofilm formation in a USA300 skin abscess isolate (UAS391) that formed prolific biofilms.
USA300 S. aureus strains, TCH1516 and FPR3757, were found to be closely related based on whole genome mapping (Argus™ Optical Mapping System, Opgen Inc, Gaithersburg, USA) to UAS391 (96.3-99.1 % similarity, P=0.0151), however differed markedly in biofilm formation (P=0.0001) on a dynamic assay (BioFlux 200, Fluxion Biosciences, USA). Comparison of whole genome sequences of these strains identified differences in a total of 18 genes. Corresponding Tn (bursa aurealis-bearing) knockout mutants in these target genes were obtained from a publicly available mutant library of the same clonal lineage (USA300-JE2) and were characterized phenotypically for biofilm formation. Tn mutants showing significant differences in biofilm formation were utilized for transduction into a plasmid-cured erythromycin-sensitive derivative of UAS391 and for complementation experiments. All strains were tested on the dynamic assay, and 17h-biofilms were stained (SYTO9, Life Technologies) and fluorescence intensity quantified by microscopy (Zeiss, ImageJ). Gene expression levels in Tn and transduced mutants were studied by quantitative reverse transcriptase PCR (StepOnePlusTM, Applied Biosystems®).
Comparison of the sequenced genomes of TCH1516, FPR3757 and UAS391 yielded a limited number of variant genes (n=18) that were hypothesized to account for the observed difference in biofilm-forming capacity. Screening of Tn mutants disrupted in these target genes identified one mutant (NE229) bearing a transposon insertion in SAUSA300_1119 (fakA), which exhibited increased biofilm formation similar to UAS391 (P=0.9320). Transduction experiments confirmed that fakA::Tn corresponded to 1.9- to 4.6-fold increase in biofilm formation depending on the USA300 strain background (P≤0.0007), while complementation of the TCH1516 wild-type fakA allele in UAS391 resulted in a 4.3-fold reduction in biofilm formation (P<0.0001).
This sequential approach, consisting of strain typing, genome comparison and functional genomics, identified fakA, a recently described fatty acid kinase in S. aureus that is essential for phospholipid synthesis and also impacts the transcription of numerous virulence factors, as a negative regulator of biofilm formation in S. aureus USA300.</abstract><cop>England</cop><pub>BioMed Central Ltd</pub><pmid>26502874</pmid><doi>10.1186/s12864-015-1956-8</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | BMC genomics, 2015-10, Vol.16 (1), p.861-861, Article 861 |
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| source | MEDLINE; Springer Online Journals Complete; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central Open Access; Springer Nature OA Free Journals; PubMed Central |
| subjects | Abscess Anopheles Bacterial Proteins - genetics Bacterial Proteins - metabolism Biofilms - growth & development Comparative analysis Genetic aspects Genome, Bacterial - genetics Genomics Growth Health aspects Mutation Plasmids - genetics Staphylococcus aureus - enzymology Staphylococcus aureus - genetics Staphylococcus aureus - physiology Transposons |
| title | Fatty acid kinase A is an important determinant of biofilm formation in Staphylococcus aureus USA300 |
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