Unveiling the substrate specificity of the ABC transporter Tba and its role in glycopeptide biosynthesis
Summary: Glycopeptide antibiotics (GPA) such as vancomycin are essential last-resort antibiotics produced by actinomycetes. Their biosynthesis is encoded within biosynthetic gene clusters, also harboring genes for regulation, and transport. Diverse types of GPAs have been characterized that differ i...
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Elsevier
2025-04-01
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004225003955 |
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| author | Nicola Gericke Dardan Beqaj Thales Kronenberger Andreas Kulik Athina Gavriilidou Mirita Franz-Wachtel Ulrich Schoppmeier Theresa Harbig Johanna Rapp Iwan Grin Nadine Ziemert Hannes Link Kay Nieselt Boris Macek Wolfgang Wohlleben Evi Stegmann Samuel Wagner |
| author_facet | Nicola Gericke Dardan Beqaj Thales Kronenberger Andreas Kulik Athina Gavriilidou Mirita Franz-Wachtel Ulrich Schoppmeier Theresa Harbig Johanna Rapp Iwan Grin Nadine Ziemert Hannes Link Kay Nieselt Boris Macek Wolfgang Wohlleben Evi Stegmann Samuel Wagner |
| author_sort | Nicola Gericke |
| collection | DOAJ |
| description | Summary: Glycopeptide antibiotics (GPA) such as vancomycin are essential last-resort antibiotics produced by actinomycetes. Their biosynthesis is encoded within biosynthetic gene clusters, also harboring genes for regulation, and transport. Diverse types of GPAs have been characterized that differ in peptide backbone composition and modification patterns. However, little is known about the ATP-binding cassette (ABC) transporters facilitating GPA export. Employing a multifaceted approach, we investigated the substrate specificity of GPA ABC-transporters toward the type-I GPA balhimycin. Phylogenetic analysis suggested and trans-complementation experiments confirmed that balhimycin is exported only by the related type I GPA transporters Tba and Tva (transporter of vancomycin). Molecular dynamics simulations and mutagenesis experiments showed that Tba exhibits specificity toward the peptide backbone rather than the modifications. Unexpectedly, deletion or functional inactivation of Tba halted balhimycin biosynthesis. Combined with proximity biotinylation experiments, this suggested that the interaction of the active transporter with the biosynthetic machinery is required for biosynthesis. |
| format | Article |
| id | doaj-art-bc43179dcb71482fa3bf8dca0655f78d |
| institution | Kabale University |
| issn | 2589-0042 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj-art-bc43179dcb71482fa3bf8dca0655f78d2025-08-20T03:42:39ZengElsevieriScience2589-00422025-04-0128411213510.1016/j.isci.2025.112135Unveiling the substrate specificity of the ABC transporter Tba and its role in glycopeptide biosynthesisNicola Gericke0Dardan Beqaj1Thales Kronenberger2Andreas Kulik3Athina Gavriilidou4Mirita Franz-Wachtel5Ulrich Schoppmeier6Theresa Harbig7Johanna Rapp8Iwan Grin9Nadine Ziemert10Hannes Link11Kay Nieselt12Boris Macek13Wolfgang Wohlleben14Evi Stegmann15Samuel Wagner16Cellular and Molecular Microbiology, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Elfriede-Aulhorn-Str. 6, 72076 Tübingen, GermanyMicrobial Active Compounds, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, GermanyCellular and Molecular Microbiology, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Elfriede-Aulhorn-Str. 6, 72076 Tübingen, Germany; Partner-Site: DZIF Tübingen, Elfriede-Aulhorn-Str. 6/Auf der Morgenstelle 28, 72076 Tübingen, Germany; School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Yliopistonrinne 3, 70211 Kuopio, FinlandMicrobial Active Compounds, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, GermanyTranslational Genome Mining for Natural Products, Interfaculty Institute of Microbiology and Infection Medicine Tübingen (IMIT), University of Tübingen, Auf der Morgenstelle 24, 72076 Tübingen, GermanyProteome Center Tübingen, Institute of Cell Biology, University of Tübingen, Auf der Morgenstelle 15, 72076 Tübingen, GermanyCellular and Molecular Microbiology, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Elfriede-Aulhorn-Str. 6, 72076 Tübingen, Germany; Excellence Cluster ''Controlling Microbes to Fight Infections'' (CMFI), University of Tübingen, 72076 Tübingen, GermanyInterfaculty Institute for Bioinformatics and Medical Informatics (IBMI), University of Tübingen, Sand 14, 72076 Tübingen, GermanyExcellence Cluster ''Controlling Microbes to Fight Infections'' (CMFI), University of Tübingen, 72076 Tübingen, Germany; Bacterial Metabolomics, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, GermanyCellular and Molecular Microbiology, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Elfriede-Aulhorn-Str. 6, 72076 Tübingen, GermanyPartner-Site: DZIF Tübingen, Elfriede-Aulhorn-Str. 6/Auf der Morgenstelle 28, 72076 Tübingen, Germany; Translational Genome Mining for Natural Products, Interfaculty Institute of Microbiology and Infection Medicine Tübingen (IMIT), University of Tübingen, Auf der Morgenstelle 24, 72076 Tübingen, GermanyExcellence Cluster ''Controlling Microbes to Fight Infections'' (CMFI), University of Tübingen, 72076 Tübingen, Germany; Bacterial Metabolomics, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, GermanyInterfaculty Institute for Bioinformatics and Medical Informatics (IBMI), University of Tübingen, Sand 14, 72076 Tübingen, GermanyProteome Center Tübingen, Institute of Cell Biology, University of Tübingen, Auf der Morgenstelle 15, 72076 Tübingen, GermanyPartner-Site: DZIF Tübingen, Elfriede-Aulhorn-Str. 6/Auf der Morgenstelle 28, 72076 Tübingen, Germany; Excellence Cluster ''Controlling Microbes to Fight Infections'' (CMFI), University of Tübingen, 72076 Tübingen, Germany; Microbiology/Biotechnology, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, GermanyMicrobial Active Compounds, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany; Partner-Site: DZIF Tübingen, Elfriede-Aulhorn-Str. 6/Auf der Morgenstelle 28, 72076 Tübingen, Germany; Excellence Cluster ''Controlling Microbes to Fight Infections'' (CMFI), University of Tübingen, 72076 Tübingen, Germany; Corresponding authorCellular and Molecular Microbiology, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Elfriede-Aulhorn-Str. 6, 72076 Tübingen, Germany; Partner-Site: DZIF Tübingen, Elfriede-Aulhorn-Str. 6/Auf der Morgenstelle 28, 72076 Tübingen, Germany; Excellence Cluster ''Controlling Microbes to Fight Infections'' (CMFI), University of Tübingen, 72076 Tübingen, Germany; Corresponding authorSummary: Glycopeptide antibiotics (GPA) such as vancomycin are essential last-resort antibiotics produced by actinomycetes. Their biosynthesis is encoded within biosynthetic gene clusters, also harboring genes for regulation, and transport. Diverse types of GPAs have been characterized that differ in peptide backbone composition and modification patterns. However, little is known about the ATP-binding cassette (ABC) transporters facilitating GPA export. Employing a multifaceted approach, we investigated the substrate specificity of GPA ABC-transporters toward the type-I GPA balhimycin. Phylogenetic analysis suggested and trans-complementation experiments confirmed that balhimycin is exported only by the related type I GPA transporters Tba and Tva (transporter of vancomycin). Molecular dynamics simulations and mutagenesis experiments showed that Tba exhibits specificity toward the peptide backbone rather than the modifications. Unexpectedly, deletion or functional inactivation of Tba halted balhimycin biosynthesis. Combined with proximity biotinylation experiments, this suggested that the interaction of the active transporter with the biosynthetic machinery is required for biosynthesis.http://www.sciencedirect.com/science/article/pii/S2589004225003955BiosynthesisBiochemistryChemical synthesis |
| spellingShingle | Nicola Gericke Dardan Beqaj Thales Kronenberger Andreas Kulik Athina Gavriilidou Mirita Franz-Wachtel Ulrich Schoppmeier Theresa Harbig Johanna Rapp Iwan Grin Nadine Ziemert Hannes Link Kay Nieselt Boris Macek Wolfgang Wohlleben Evi Stegmann Samuel Wagner Unveiling the substrate specificity of the ABC transporter Tba and its role in glycopeptide biosynthesis iScience Biosynthesis Biochemistry Chemical synthesis |
| title | Unveiling the substrate specificity of the ABC transporter Tba and its role in glycopeptide biosynthesis |
| title_full | Unveiling the substrate specificity of the ABC transporter Tba and its role in glycopeptide biosynthesis |
| title_fullStr | Unveiling the substrate specificity of the ABC transporter Tba and its role in glycopeptide biosynthesis |
| title_full_unstemmed | Unveiling the substrate specificity of the ABC transporter Tba and its role in glycopeptide biosynthesis |
| title_short | Unveiling the substrate specificity of the ABC transporter Tba and its role in glycopeptide biosynthesis |
| title_sort | unveiling the substrate specificity of the abc transporter tba and its role in glycopeptide biosynthesis |
| topic | Biosynthesis Biochemistry Chemical synthesis |
| url | http://www.sciencedirect.com/science/article/pii/S2589004225003955 |
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