Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli
Cell-penetrating peptides (CPPs) are increasingly used for cellular drug delivery in both pro- and eukaryotic cells, and oligoarginines have attracted special attention. How arginine-rich CPPs translocate across the cell envelope, particularly for prokaryotes, is still unknown. Arginine-rich CPPs ef...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-09-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253121001505 |
| _version_ | 1819138149596528640 |
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| author | Jakob Frimodt-Møller Andreas Koulouktsis Godefroid Charbon Marit Otterlei Peter E. Nielsen Anders Løbner-Olesen |
| author_facet | Jakob Frimodt-Møller Andreas Koulouktsis Godefroid Charbon Marit Otterlei Peter E. Nielsen Anders Løbner-Olesen |
| author_sort | Jakob Frimodt-Møller |
| collection | DOAJ |
| description | Cell-penetrating peptides (CPPs) are increasingly used for cellular drug delivery in both pro- and eukaryotic cells, and oligoarginines have attracted special attention. How arginine-rich CPPs translocate across the cell envelope, particularly for prokaryotes, is still unknown. Arginine-rich CPPs efficiently deliver antimicrobial peptide nucleic acid (PNA) to its intracellular mRNA target in bacteria. We show that resistance to PNA conjugated to an arginine-rich CPP in Escherichia coli requires multiple genetic modifications and is specific for the CPP part and not to the PNA part. An integral part of the resistance was the constitutively activated Cpx-envelope stress response system (cpx∗), which decreased the cytoplasmic membrane potential. This indicates an indirect energy-dependent uptake mechanism for antimicrobials conjugated to arginine-rich CPPs. In agreement, cpx∗ mutants showed low-level resistance to aminoglycosides and an arginine-rich CPP conjugated to a peptide targeting the DNA sliding clamp, i.e., similar uptake in E. coli for these antimicrobial compounds. |
| first_indexed | 2024-12-22T11:02:11Z |
| format | Article |
| id | doaj.art-253da9a6a4314fa899fff9d789465764 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-22T11:02:11Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-253da9a6a4314fa899fff9d7894657642022-12-21T18:28:28ZengElsevierMolecular Therapy: Nucleic Acids2162-25312021-09-0125444454Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coliJakob Frimodt-Møller0Andreas Koulouktsis1Godefroid Charbon2Marit Otterlei3Peter E. Nielsen4Anders Løbner-Olesen5Department of Biology, Center for Peptide-Based Antibiotics, University of Copenhagen, Ole Maaløes vej 5, 2200 Copenhagen N, Denmark; Corresponding author: Jakob Frimodt-Møller, Department of Biology, Center for Peptide-Based Antibiotics, University of Copenhagen, Ole Maaløes vej 5, 2200 Copenhagen N, Denmark.Department of Biology, Center for Peptide-Based Antibiotics, University of Copenhagen, Ole Maaløes vej 5, 2200 Copenhagen N, DenmarkDepartment of Biology, Center for Peptide-Based Antibiotics, University of Copenhagen, Ole Maaløes vej 5, 2200 Copenhagen N, DenmarkDepartment of Clinical and Molecular Medicine, Faculty of Medicine and Health Sciences, NTNU, Norwegian University of Science and Technology, NO-7489 Trondheim, NorwayDepartment of Cellular and Molecular Medicine, Center for Peptide-Based Antibiotics, The Panum Institute, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen N, DenmarkDepartment of Biology, Center for Peptide-Based Antibiotics, University of Copenhagen, Ole Maaløes vej 5, 2200 Copenhagen N, Denmark; Corresponding author: Anders Løbner-Olesen, Department of Biology, Center for Peptide-Based Antibiotics, University of Copenhagen, Ole Maaløes vej 5, 2200 Copenhagen N, Denmark.Cell-penetrating peptides (CPPs) are increasingly used for cellular drug delivery in both pro- and eukaryotic cells, and oligoarginines have attracted special attention. How arginine-rich CPPs translocate across the cell envelope, particularly for prokaryotes, is still unknown. Arginine-rich CPPs efficiently deliver antimicrobial peptide nucleic acid (PNA) to its intracellular mRNA target in bacteria. We show that resistance to PNA conjugated to an arginine-rich CPP in Escherichia coli requires multiple genetic modifications and is specific for the CPP part and not to the PNA part. An integral part of the resistance was the constitutively activated Cpx-envelope stress response system (cpx∗), which decreased the cytoplasmic membrane potential. This indicates an indirect energy-dependent uptake mechanism for antimicrobials conjugated to arginine-rich CPPs. In agreement, cpx∗ mutants showed low-level resistance to aminoglycosides and an arginine-rich CPP conjugated to a peptide targeting the DNA sliding clamp, i.e., similar uptake in E. coli for these antimicrobial compounds.http://www.sciencedirect.com/science/article/pii/S2162253121001505peptide nucleic acidsCpx responseaminoglycosidesarginine-rich cell-penetrating peptidesinner membrane |
| spellingShingle | Jakob Frimodt-Møller Andreas Koulouktsis Godefroid Charbon Marit Otterlei Peter E. Nielsen Anders Løbner-Olesen Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli Molecular Therapy: Nucleic Acids peptide nucleic acids Cpx response aminoglycosides arginine-rich cell-penetrating peptides inner membrane |
| title | Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli |
| title_full | Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli |
| title_fullStr | Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli |
| title_full_unstemmed | Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli |
| title_short | Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli |
| title_sort | activating the cpx response induces tolerance to antisense pna delivered by an arginine rich peptide in escherichia coli |
| topic | peptide nucleic acids Cpx response aminoglycosides arginine-rich cell-penetrating peptides inner membrane |
| url | http://www.sciencedirect.com/science/article/pii/S2162253121001505 |
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