Multiple routes for non-physiological l-threonine uptake in Escherichia coli K-12
In this study, we identified eight multicopy suppressors (yhjE, sdaC, ydgI, alaE, ychE, yqeG, proP, and yjeM) and three distinct classes of chromosomal mutations (lrp, marC, and cycA) capable of complementing the growth defect caused by threonine uptake deficiency in the sstT tdcC livKHMGF brnQ thrP...
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Frontiers Media S.A.
2025-04-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1579813/full |
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| author | Dmitrii M. Bubnov Andrey A. Khozov Andrey A. Khozov Tatiana V. Vybornaya Agnessa A. Stepanova Agnessa A. Stepanova Sergey V. Molev Sergey V. Molev Olga E. Melkina Gennadii A. Badun Maria G. Chernysheva Ilia A. Skob Alexander I. Netrusov Sergey P. Sineoky |
| author_facet | Dmitrii M. Bubnov Andrey A. Khozov Andrey A. Khozov Tatiana V. Vybornaya Agnessa A. Stepanova Agnessa A. Stepanova Sergey V. Molev Sergey V. Molev Olga E. Melkina Gennadii A. Badun Maria G. Chernysheva Ilia A. Skob Alexander I. Netrusov Sergey P. Sineoky |
| author_sort | Dmitrii M. Bubnov |
| collection | DOAJ |
| description | In this study, we identified eight multicopy suppressors (yhjE, sdaC, ydgI, alaE, ychE, yqeG, proP, and yjeM) and three distinct classes of chromosomal mutations (lrp, marC, and cycA) capable of complementing the growth defect caused by threonine uptake deficiency in the sstT tdcC livKHMGF brnQ thrP strain. YhjE, SdaC, YdgI, AlaE, mutant MarC, and CycA exhibited measurable threonine-specific uptake activity in the in vitro assay. Phenotypic assays revealed that YhjE and SdaC were the main entry points for threonine in a strain lacking major threonine-specific permeases. A derivative of the threonine-auxotrophic sstT tdcC livKHMGF brnQ thrP mutant, harboring deletions of eight multicopy suppressors, exhibited significantly reduced fitness at subsaturating threonine concentrations and improved fitness at toxic threonine concentrations, indicating a defect in membrane permeability. These results may help guide the effective construction of threonine-producing strains, extend knowledge on the substrate preferences of SdaC, AlaE, and ProP, and provide clues for further studies on the exact substrate range of YhjE, YdgI, YjeM, YchE, MarC, and YqeG whose physiologically relevant functions have not yet been established. |
| format | Article |
| id | doaj-art-da59991387ae4d779a94a4d90d1272fc |
| institution | DOAJ |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj-art-da59991387ae4d779a94a4d90d1272fc2025-08-20T03:06:43ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-04-011610.3389/fmicb.2025.15798131579813Multiple routes for non-physiological l-threonine uptake in Escherichia coli K-12Dmitrii M. Bubnov0Andrey A. Khozov1Andrey A. Khozov2Tatiana V. Vybornaya3Agnessa A. Stepanova4Agnessa A. Stepanova5Sergey V. Molev6Sergey V. Molev7Olga E. Melkina8Gennadii A. Badun9Maria G. Chernysheva10Ilia A. Skob11Alexander I. Netrusov12Sergey P. Sineoky13National Research Centre “Kurchatov Institute”, Moscow, RussiaNational Research Centre “Kurchatov Institute”, Moscow, RussiaDepartment of Microbiology, Faculty of Biology, Lomonosov Moscow State University, Moscow, RussiaNational Research Centre “Kurchatov Institute”, Moscow, RussiaNational Research Centre “Kurchatov Institute”, Moscow, RussiaDepartment of Microbiology, Faculty of Biology, Lomonosov Moscow State University, Moscow, RussiaNational Research Centre “Kurchatov Institute”, Moscow, RussiaDepartment of Microbiology, Faculty of Biology, Lomonosov Moscow State University, Moscow, RussiaNational Research Centre “Kurchatov Institute”, Moscow, RussiaDepartment of Radiochemistry, Faculty of Chemistry, Lomonosov Moscow State University, Moscow, RussiaDepartment of Radiochemistry, Faculty of Chemistry, Lomonosov Moscow State University, Moscow, RussiaDepartment of Genetics, Faculty of Biology, Lomonosov Moscow State University, Moscow, RussiaDepartment of Microbiology, Faculty of Biology, Lomonosov Moscow State University, Moscow, RussiaNational Research Centre “Kurchatov Institute”, Moscow, RussiaIn this study, we identified eight multicopy suppressors (yhjE, sdaC, ydgI, alaE, ychE, yqeG, proP, and yjeM) and three distinct classes of chromosomal mutations (lrp, marC, and cycA) capable of complementing the growth defect caused by threonine uptake deficiency in the sstT tdcC livKHMGF brnQ thrP strain. YhjE, SdaC, YdgI, AlaE, mutant MarC, and CycA exhibited measurable threonine-specific uptake activity in the in vitro assay. Phenotypic assays revealed that YhjE and SdaC were the main entry points for threonine in a strain lacking major threonine-specific permeases. A derivative of the threonine-auxotrophic sstT tdcC livKHMGF brnQ thrP mutant, harboring deletions of eight multicopy suppressors, exhibited significantly reduced fitness at subsaturating threonine concentrations and improved fitness at toxic threonine concentrations, indicating a defect in membrane permeability. These results may help guide the effective construction of threonine-producing strains, extend knowledge on the substrate preferences of SdaC, AlaE, and ProP, and provide clues for further studies on the exact substrate range of YhjE, YdgI, YjeM, YchE, MarC, and YqeG whose physiologically relevant functions have not yet been established.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1579813/fullEscherichia colil-threonine uptaketransmembrane transportamino acid transportermembrane proteins |
| spellingShingle | Dmitrii M. Bubnov Andrey A. Khozov Andrey A. Khozov Tatiana V. Vybornaya Agnessa A. Stepanova Agnessa A. Stepanova Sergey V. Molev Sergey V. Molev Olga E. Melkina Gennadii A. Badun Maria G. Chernysheva Ilia A. Skob Alexander I. Netrusov Sergey P. Sineoky Multiple routes for non-physiological l-threonine uptake in Escherichia coli K-12 Frontiers in Microbiology Escherichia coli l-threonine uptake transmembrane transport amino acid transporter membrane proteins |
| title | Multiple routes for non-physiological l-threonine uptake in Escherichia coli K-12 |
| title_full | Multiple routes for non-physiological l-threonine uptake in Escherichia coli K-12 |
| title_fullStr | Multiple routes for non-physiological l-threonine uptake in Escherichia coli K-12 |
| title_full_unstemmed | Multiple routes for non-physiological l-threonine uptake in Escherichia coli K-12 |
| title_short | Multiple routes for non-physiological l-threonine uptake in Escherichia coli K-12 |
| title_sort | multiple routes for non physiological l threonine uptake in escherichia coli k 12 |
| topic | Escherichia coli l-threonine uptake transmembrane transport amino acid transporter membrane proteins |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1579813/full |
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