Cell Wall Antibiotics Provoke Accumulation of Anchored mCherry in the Cross Wall of Staphylococcus aureus
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{"title"=>"Cell wall antibiotics provoke accumulation of anchored mcherry in the cross wall of staphylococcus aureus", "type"=>"journal", "authors"=>[{"first_name"=>"Wenqi", "last_name"=>"Yu", "scopus_author_id"=>"35794712100"}, {"first_name"=>"Friedrich", "last_name"=>"Götz", "scopus_author_id"=>"7102980726"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"22253886", "sgr"=>"84855600451", "doi"=>"10.1371/journal.pone.0030076", "scopus"=>"2-s2.0-84855600451", "pui"=>"364062216", "isbn"=>"1932-6203", "issn"=>"19326203"}, "id"=>"69fc806e-3b06-3842-bf28-c21b670b2fbf", "abstract"=>"A fluorescence microscopy method to directly follow the localization of defined proteins in Staphylococcus was hampered by the unstable fluorescence of fluorescent proteins. Here, we constructed plasmid (pCX) encoded red fluorescence (RF) mCherry (mCh) hybrids, namely mCh-cyto (no signal peptide and no sorting sequence), mCh-sec (with signal peptide), and mCh-cw (with signal peptide and cell wall sorting sequence). The S. aureus clones targeted mCh-fusion proteins into the cytosol, the supernatant and the cell envelope respectively; in all cases mCherry exhibited bright fluorescence. In staphylococci two types of signal peptides (SP) can be distinguished: the +YSIRK motif SP(lip) and the -YSIRK motif SP(sasF). mCh-hybrids supplied with the +YSIRK motif SP(lip) were always expressed higher than those with -YSIRK motif SP(sasF). To study the location of the anchoring process and also the influence of SP type, mCh-cw was supplied on the one hand with +YSIRK motif (mCh-cw1) and the other hand with -YSIRK motif (mCh-cw2). MCh-cw1 preferentially localized at the cross wall, while mCh-cw2 preferentially localized at the peripheral wall. Interestingly, when treated with sub-lethal concentrations of penicillin or moenomycin, both mCh-cw1 and mCh-cw2 were concentrated at the cross wall. The shift from the peripheral wall to the cross wall required Sortase A (SrtA), as in the srtA mutant this effect was blunted. The effect is most likely due to antibiotic mediated increase of free anchoring sites (Lipid II) at the cross wall, the substrate of SrtA, leading to a preferential incorporation of anchored proteins at the cross wall.", "link"=>"http://www.mendeley.com/research/cell-wall-antibiotics-provoke-accumulation-anchored-mcherry-cross-wall-staphylococcus-aureus", "reader_count"=>28, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Student > Doctoral Student"=>3, "Researcher"=>11, "Student > Ph. D. Student"=>6, "Student > Master"=>1, "Other"=>2, "Student > Bachelor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Student > Doctoral Student"=>3, "Researcher"=>11, "Student > Ph. D. Student"=>6, "Student > Master"=>1, "Other"=>2, "Student > Bachelor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>17, "Medicine and Dentistry"=>1, "Design"=>1, "Chemistry"=>3, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Design"=>{"Design"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>3}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>17}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>3}}, "reader_count_by_country"=>{"Belgium"=>1, "France"=>1, "Switzerland"=>1, "Portugal"=>1, "Germany"=>2}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/353104", "https://ndownloader.figshare.com/files/353158", "https://ndownloader.figshare.com/files/353191"], "description"=>"<div><p>A fluorescence microscopy method to directly follow the localization of defined proteins in <em>Staphylococcus</em> was hampered by the unstable fluorescence of fluorescent proteins. Here, we constructed plasmid (pCX) encoded red fluorescence (RF) mCherry (mCh) hybrids, namely mCh-cyto (no signal peptide and no sorting sequence), mCh-sec (with signal peptide), and mCh-cw (with signal peptide and cell wall sorting sequence). The <em>S. aureus</em> clones targeted mCh-fusion proteins into the cytosol, the supernatant and the cell envelope respectively; in all cases mCherry exhibited bright fluorescence. In staphylococci two types of signal peptides (SP) can be distinguished: the +YSIRK motif SP<sub>lip</sub> and the −YSIRK motif SP<sub>sasF</sub>. mCh-hybrids supplied with the +YSIRK motif SP<sub>lip</sub> were always expressed higher than those with −YSIRK motif SP<sub>sasF</sub>. To study the location of the anchoring process and also the influence of SP type, mCh-cw was supplied on the one hand with +YSIRK motif (mCh-cw1) and the other hand with -YSIRK motif (mCh-cw2). MCh-cw1 preferentially localized at the cross wall, while mCh-cw2 preferentially localized at the peripheral wall. Interestingly, when treated with sub-lethal concentrations of penicillin or moenomycin, both mCh-cw1 and mCh-cw2 were concentrated at the cross wall. The shift from the peripheral wall to the cross wall required Sortase A (SrtA), as in the <em>srtA</em> mutant this effect was blunted. The effect is most likely due to antibiotic mediated increase of free anchoring sites (Lipid II) at the cross wall, the substrate of SrtA, leading to a preferential incorporation of anchored proteins at the cross wall.</p> </div>", "links"=>[], "tags"=>["antibiotics", "accumulation", "anchored", "mcherry"], "article_id"=>129801, "categories"=>["Biochemistry", "Microbiology"], "users"=>["Wenqi Yu", "Friedrich Götz"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030076.s001", "https://dx.doi.org/10.1371/journal.pone.0030076.s002", "https://dx.doi.org/10.1371/journal.pone.0030076.s003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Cell_Wall_Antibiotics_Provoke_Accumulation_of_Anchored_mCherry_in_the_Cross_Wall_of_Staphylococcus_aureus_/129801", "title"=>"Cell Wall Antibiotics Provoke Accumulation of Anchored mCherry in the Cross Wall of <em>Staphylococcus aureus</em>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-01-10 02:43:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/693985"], "description"=>"<p>SP, signal peptide; PP, propeptide; CWS, cell wall sorting signal; mCh: mCherry; lip, lipase. The amino acid sequence of CWS was indicated. The parent plasmid was pCX30 and all mCh-fusion constructs were under control of the xylose-inducible promoter, P<i>xyl</i>.</p>", "links"=>[], "tags"=>["microbiology", "Biochemistry"], "article_id"=>364428, "categories"=>["Biochemistry", "Microbiology"], "users"=>["Wenqi Yu", "Friedrich Götz"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030076.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_representation_of_mCh_hybrids_/364428", "title"=>"Schematic representation of mCh-hybrids.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-10 01:13:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/694067"], "description"=>"<p><b>A</b>. Fluorescence intensity comparison of mCh-hybrids from different cell fractions. WT-cyto, SA113 (pCXmCh-cyto); WT-cw1 or 2, SA113 (pCXmCh-cw1) or (pCXmCh-cw2); WT-sec1 or 2, SA113 (pCXmCh-sec1) or (pCXmCh-sec2); Δ<i>srtA</i>-cw1 or 2, SA113 Δ<i>srtA</i> (pCXmCh-cw1) or (pCXmCh-cw2); lys, lysostaphin. <b>B.</b> Western blotting of mCh-hybrid proteins in the culture supernatant of protein A deficient mutant SA113 Δ<i>spa</i>. Blank, SA113 Δ<i>spa</i> without plasmid; cyto, SA113 Δ<i>spa</i> (pCXmCh-cyto); cw1 or 2, SA113 Δ<i>spa</i> (pCXmCh-cw1) or (pCXmCh-cw2); sec1 or 2, SA113 Δ<i>spa</i> (pCXmCh-sec1) or (pCXmCh-sec2); Δ<i>srtA</i>-cw1 or 2, SA113 Δ<i>spa</i>Δ<i>srtA</i> (pCXmCh-cw1) or (pCXmCh-cw2). <b>C.</b> Subcellular localization of mCh-hybrid proteins in SA113. <b>a.</b> pCXmCh-cw1; <b>b.</b> pCXmCh-cw2; <b>c.</b> pCXmCh-sec1; <b>d.</b> pCXmCh-sec2; <b>e.</b> pCXmCh-cyto. Arrowheads in <b>a</b> and <b>b</b>, fluorescence localized at the cross wall in <b>a</b>, but absent from the cross wall in <b>b</b>; arrows in <b>a</b> and <b>c,</b> RF foci close to the initial sites of the cross walls; arrowheads in <b>d,</b> halo-like RF distribution absent from the cross wall. Images <b>a, c,</b> and <b>e</b> were taken after one hour of xylose induction; images <b>b</b> and <b>d</b> were taken after two hours of induction. Green: Van-FL staining (cell wall); scale bar, 2 µm.</p>", "links"=>[], "tags"=>["microbiology", "Biochemistry"], "article_id"=>364513, "categories"=>["Biochemistry", "Microbiology"], "users"=>["Wenqi Yu", "Friedrich Götz"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030076.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Monitoring_mCh_hybrids_/364513", "title"=>"Monitoring mCh-hybrids.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-10 01:15:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/694216"], "description"=>"<p><b>A.</b> Schematic representation of antibiotics treatment assay. Untreated (<b>□</b>); treated with penicillin (0.02 µg/ml) (•); treated with moenomycin (flavomycin) (1 µg/ml) (×). <b>B.</b> Influence of penicillin (Pc) and moenomycin (Fla) on the subcellular localization of mCh-cw hybrid proteins. Arrowheads indicated the cross wall accumulation of mCh-cw; arrows indicated the ring-like distribution; scale bar, 2 µm.</p>", "links"=>[], "tags"=>["moenomycin", "mch-cw", "irrespective", "sp"], "article_id"=>364652, "categories"=>["Biochemistry", "Microbiology"], "users"=>["Wenqi Yu", "Friedrich Götz"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030076.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Penicillin_and_moenomycin_direct_mCh_cw_to_the_cross_wall_irrespective_of_SP_type_/364652", "title"=>"Penicillin and moenomycin direct mCh-cw to the cross wall, irrespective of SP type.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-10 01:17:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/694329"], "description"=>"<p><b>A.</b> Fluorescence intensity profile of Van-FL staining from a line perpendicular to the cross wall and across the middle point of the cross wall. Simple line, untreated cell; dotted line with filled squares, moenomycin (Fla) treated cell; line with filled circles, penicillin (Pc) treated cell. Max amplitude represented the cross wall intensity. Note that the figure was remade using ImageJ software from the microscopy images; the intensity and distance values were not the same as the original data from Leica AF software; but represented the same profile distribution. <b>B.</b> Comparative Van-FL intensity at the cross wall among untreated, penicillin (Pc) treated, and moenomycin (Fla) treated cells. The cross wall Van-FL intensity values were calculated by the ratio of max amplitude against mean fluorescence intensity (generated by Leica AF software) from the same cell. The average ratio of 150 cells from three independent experiments of each group was shown in the bars. White bar, SA113 (pCXmCh-cw1); gray bar, SA113 (pCXmCh-cw2).</p>", "links"=>[], "tags"=>["moenomycin", "led", "enrichment", "van-fl"], "article_id"=>364764, "categories"=>["Biochemistry", "Microbiology"], "users"=>["Wenqi Yu", "Friedrich Götz"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030076.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Penicillin_and_moenomycin_treatment_led_to_enrichment_of_Van_FL_at_the_cross_wall_/364764", "title"=>"Penicillin and moenomycin treatment led to enrichment of Van-FL at the cross wall.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-10 01:19:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/694433"], "description"=>"<p>Arrows, mCh-cw dispersed over the entire cell; arrowheads, the cross wall localized mCh-cw. Scale bar, 2 µm.</p>", "links"=>[], "tags"=>["patterns", "penicillin"], "article_id"=>364868, "categories"=>["Biochemistry", "Microbiology"], "users"=>["Wenqi Yu", "Friedrich Götz"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030076.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Localization_patterns_of_srtA_pCXmCh_cw1_2_in_the_presence_of_penicillin_or_moenomycin_/364868", "title"=>"Localization patterns of Δ<i>srtA</i> (pCXmCh-cw1&2) in the presence of penicillin or moenomycin.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-10 01:21:08"}

PMC Usage Stats | Further Information

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Relative Metric

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