Antibacterial potency of type VI amidase effector toxins is dependent on substrate topology and cellular context
Members of the bacterial T6SS amidase effector (Tae) superfamily of toxins are delivered between competing bacteria to degrade cell wall peptidoglycan. Although Taes share a common substrate, they exhibit distinct antimicrobial potency across different competitor species. To investigate the molecula...
| Main Authors: | , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-06-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/79796 |
| _version_ | 1818018357328216064 |
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| author | Atanas Radkov Anne L Sapiro Sebastian Flores Corey Henderson Hayden Saunders Rachel Kim Steven Massa Samuel Thompson Chase Mateusiak Jacob Biboy Ziyi Zhao Lea M Starita William L Hatleberg Waldemar Vollmer Alistair B Russell Jean-Pierre Simorre Spencer Anthony-Cahill Peter Brzovic Beth Hayes Seemay Chou |
| author_facet | Atanas Radkov Anne L Sapiro Sebastian Flores Corey Henderson Hayden Saunders Rachel Kim Steven Massa Samuel Thompson Chase Mateusiak Jacob Biboy Ziyi Zhao Lea M Starita William L Hatleberg Waldemar Vollmer Alistair B Russell Jean-Pierre Simorre Spencer Anthony-Cahill Peter Brzovic Beth Hayes Seemay Chou |
| author_sort | Atanas Radkov |
| collection | DOAJ |
| description | Members of the bacterial T6SS amidase effector (Tae) superfamily of toxins are delivered between competing bacteria to degrade cell wall peptidoglycan. Although Taes share a common substrate, they exhibit distinct antimicrobial potency across different competitor species. To investigate the molecular basis governing these differences, we quantitatively defined the functional determinants of Tae1 from Pseudomonas aeruginosa PAO1 using a combination of nuclear magnetic resonance and a high-throughput in vivo genetic approach called deep mutational scanning (DMS). As expected, combined analyses confirmed the role of critical residues near the Tae1 catalytic center. Unexpectedly, DMS revealed substantial contributions to enzymatic activity from a much larger, ring-like functional hot spot extending around the entire circumference of the enzyme. Comparative DMS across distinct growth conditions highlighted how functional contribution of different surfaces is highly context-dependent, varying alongside composition of targeted cell walls. These observations suggest that Tae1 engages with the intact cell wall network through a more distributed three-dimensional interaction interface than previously appreciated, providing an explanation for observed differences in antimicrobial potency across divergent Gram-negative competitors. Further binding studies of several Tae1 variants with their cognate immunity protein demonstrate that requirements to maintain protection from Tae activity may be a significant constraint on the mutational landscape of tae1 toxicity in the wild. In total, our work reveals that Tae diversification has likely been shaped by multiple independent pressures to maintain interactions with binding partners that vary across bacterial species and conditions. |
| first_indexed | 2024-04-14T07:38:04Z |
| format | Article |
| id | doaj.art-e8b6e88e62cc4ba1b7e72be730157666 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-14T07:38:04Z |
| publishDate | 2022-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-e8b6e88e62cc4ba1b7e72be7301576662022-12-22T02:05:37ZengeLife Sciences Publications LtdeLife2050-084X2022-06-011110.7554/eLife.79796Antibacterial potency of type VI amidase effector toxins is dependent on substrate topology and cellular contextAtanas Radkov0https://orcid.org/0000-0001-7418-279XAnne L Sapiro1https://orcid.org/0000-0002-6612-8272Sebastian Flores2Corey Henderson3Hayden Saunders4https://orcid.org/0000-0002-7582-3031Rachel Kim5https://orcid.org/0000-0003-3793-4264Steven Massa6Samuel Thompson7Chase Mateusiak8https://orcid.org/0000-0002-2890-4242Jacob Biboy9https://orcid.org/0000-0002-1286-6851Ziyi Zhao10Lea M Starita11William L Hatleberg12https://orcid.org/0000-0002-0423-7123Waldemar Vollmer13Alistair B Russell14Jean-Pierre Simorre15https://orcid.org/0000-0002-7943-1342Spencer Anthony-Cahill16Peter Brzovic17Beth Hayes18https://orcid.org/0000-0001-6633-751XSeemay Chou19https://orcid.org/0000-0002-7271-303XDepartment of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United StatesDepartment of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United StatesUniversity of Miami, Miami, United StatesInBios International, Seattle, United StatesDepartment of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United StatesPacific Northwest University of Health Sciences, Yakima, United StatesDepartment of Biology, Stanford University, Stanford, United StatesDepartment of Bioengineering, Stanford University, Stanford, United StatesComputer Science Department, Washington University in St. Louis, St. Louis, United StatesCentre for Bacterial Cell Biology, Newcastle University, Newcastle upon Tyne, United KingdomDepartment of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United StatesDepartment of Genome Sciences, University of Washington, Seattle, United States; Brotman Baty Institute for Precision Medicine, Seattle, United StatesIndependent Researcher, Pittsburg, United StatesCentre for Bacterial Cell Biology, Newcastle University, Newcastle upon Tyne, United KingdomDivision of Biological Sciences, University of California, San Diego, La Jolla, United StatesUniversité Grenoble Alpes, Grenoble, FranceChemistry Department, Western Washington University, Bellingham, United StatesDepartment of Biochemistry, University of Washington, Seattle, United StatesDepartment of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United StatesDepartment of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United StatesMembers of the bacterial T6SS amidase effector (Tae) superfamily of toxins are delivered between competing bacteria to degrade cell wall peptidoglycan. Although Taes share a common substrate, they exhibit distinct antimicrobial potency across different competitor species. To investigate the molecular basis governing these differences, we quantitatively defined the functional determinants of Tae1 from Pseudomonas aeruginosa PAO1 using a combination of nuclear magnetic resonance and a high-throughput in vivo genetic approach called deep mutational scanning (DMS). As expected, combined analyses confirmed the role of critical residues near the Tae1 catalytic center. Unexpectedly, DMS revealed substantial contributions to enzymatic activity from a much larger, ring-like functional hot spot extending around the entire circumference of the enzyme. Comparative DMS across distinct growth conditions highlighted how functional contribution of different surfaces is highly context-dependent, varying alongside composition of targeted cell walls. These observations suggest that Tae1 engages with the intact cell wall network through a more distributed three-dimensional interaction interface than previously appreciated, providing an explanation for observed differences in antimicrobial potency across divergent Gram-negative competitors. Further binding studies of several Tae1 variants with their cognate immunity protein demonstrate that requirements to maintain protection from Tae activity may be a significant constraint on the mutational landscape of tae1 toxicity in the wild. In total, our work reveals that Tae diversification has likely been shaped by multiple independent pressures to maintain interactions with binding partners that vary across bacterial species and conditions.https://elifesciences.org/articles/79796type VI amidase effectorpeptidoglycantopologycognate immunitydeep mutational scanninghost–microbe interaction |
| spellingShingle | Atanas Radkov Anne L Sapiro Sebastian Flores Corey Henderson Hayden Saunders Rachel Kim Steven Massa Samuel Thompson Chase Mateusiak Jacob Biboy Ziyi Zhao Lea M Starita William L Hatleberg Waldemar Vollmer Alistair B Russell Jean-Pierre Simorre Spencer Anthony-Cahill Peter Brzovic Beth Hayes Seemay Chou Antibacterial potency of type VI amidase effector toxins is dependent on substrate topology and cellular context eLife type VI amidase effector peptidoglycan topology cognate immunity deep mutational scanning host–microbe interaction |
| title | Antibacterial potency of type VI amidase effector toxins is dependent on substrate topology and cellular context |
| title_full | Antibacterial potency of type VI amidase effector toxins is dependent on substrate topology and cellular context |
| title_fullStr | Antibacterial potency of type VI amidase effector toxins is dependent on substrate topology and cellular context |
| title_full_unstemmed | Antibacterial potency of type VI amidase effector toxins is dependent on substrate topology and cellular context |
| title_short | Antibacterial potency of type VI amidase effector toxins is dependent on substrate topology and cellular context |
| title_sort | antibacterial potency of type vi amidase effector toxins is dependent on substrate topology and cellular context |
| topic | type VI amidase effector peptidoglycan topology cognate immunity deep mutational scanning host–microbe interaction |
| url | https://elifesciences.org/articles/79796 |
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