High Levels of Cyclic Diguanylate Interfere with Beneficial Bacterial Colonization
ABSTRACT During colonization of the Hawaiian bobtail squid (Euprymna scolopes), Vibrio fischeri bacteria undergo a lifestyle transition from a planktonic motile state in the environment to a biofilm state in host mucus. Cyclic diguanylate (c-di-GMP) is a cytoplasmic signaling molecule that is import...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2022-08-01
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| Series: | mBio |
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| Online Access: | https://journals.asm.org/doi/10.1128/mbio.01671-22 |
| _version_ | 1828743486488182784 |
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| author | Ruth Y. Isenberg David G. Christensen Karen L. Visick Mark J. Mandel |
| author_facet | Ruth Y. Isenberg David G. Christensen Karen L. Visick Mark J. Mandel |
| author_sort | Ruth Y. Isenberg |
| collection | DOAJ |
| description | ABSTRACT During colonization of the Hawaiian bobtail squid (Euprymna scolopes), Vibrio fischeri bacteria undergo a lifestyle transition from a planktonic motile state in the environment to a biofilm state in host mucus. Cyclic diguanylate (c-di-GMP) is a cytoplasmic signaling molecule that is important for regulating motility-biofilm transitions in many bacterial species. V. fischeri encodes 50 proteins predicted to synthesize and/or degrade c-di-GMP, but a role for c-di-GMP regulation during host colonization has not been investigated. We examined strains exhibiting either low or high levels of c-di-GMP during squid colonization and found that while a low-c-di-GMP strain had no colonization defect, a high c-di-GMP strain was severely impaired. Expression of a heterologous c-di-GMP phosphodiesterase restored colonization, demonstrating that the effect is due to high c-di-GMP levels. In the constitutive high-c-di-GMP state, colonizing V. fischeri exhibited reduced motility, altered biofilm aggregate morphology, and a regulatory interaction where transcription of one polysaccharide locus is inhibited by the presence of the other polysaccharide. Our results highlight the importance of proper c-di-GMP regulation during beneficial animal colonization, illustrate multiple pathways regulated by c-di-GMP in the host, and uncover an interplay of multiple exopolysaccharide systems in host-associated aggregates. IMPORTANCE There is substantial interest in studying cyclic diguanylate (c-di-GMP) in pathogenic and environmental bacteria, which has led to an accepted paradigm in which high c-di-GMP levels promote biofilm formation and reduce motility. However, considerably less focus has been placed on understanding how this compound contributes to beneficial colonization. Using the Vibrio fischeri-Hawaiian bobtail squid study system, we took advantage of recent genetic advances in the bacterium to modulate c-di-GMP levels and measure colonization and track c-di-GMP phenotypes in a symbiotic interaction. Studies in the animal host revealed a c-di-GMP-dependent genetic interaction between two distinct biofilm polysaccharides, Syp and cellulose, that was not evident in culture-based studies: elevated c-di-GMP altered the composition and abundance of the in vivo biofilm by decreasing syp transcription due to increased cellulose synthesis. This study reveals important parallels between pathogenic and beneficial colonization and additionally identifies c-di-GMP-dependent regulation that occurs specifically in the squid host. |
| first_indexed | 2024-04-13T01:39:21Z |
| format | Article |
| id | doaj.art-08bc1b86f8d04552a8678c028f7eff18 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-04-13T01:39:21Z |
| publishDate | 2022-08-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-08bc1b86f8d04552a8678c028f7eff182022-12-22T03:08:15ZengAmerican Society for MicrobiologymBio2150-75112022-08-0113410.1128/mbio.01671-22High Levels of Cyclic Diguanylate Interfere with Beneficial Bacterial ColonizationRuth Y. Isenberg0David G. Christensen1Karen L. Visick2Mark J. Mandel3Department of Medical Microbiology and Immunology, University of Wisconsin—Madison, Madison, Wisconsin, USADepartment of Microbiology and Immunology, Loyola University Stritch School of Medicine, Maywood, Illinois, USADepartment of Microbiology and Immunology, Loyola University Stritch School of Medicine, Maywood, Illinois, USADepartment of Medical Microbiology and Immunology, University of Wisconsin—Madison, Madison, Wisconsin, USAABSTRACT During colonization of the Hawaiian bobtail squid (Euprymna scolopes), Vibrio fischeri bacteria undergo a lifestyle transition from a planktonic motile state in the environment to a biofilm state in host mucus. Cyclic diguanylate (c-di-GMP) is a cytoplasmic signaling molecule that is important for regulating motility-biofilm transitions in many bacterial species. V. fischeri encodes 50 proteins predicted to synthesize and/or degrade c-di-GMP, but a role for c-di-GMP regulation during host colonization has not been investigated. We examined strains exhibiting either low or high levels of c-di-GMP during squid colonization and found that while a low-c-di-GMP strain had no colonization defect, a high c-di-GMP strain was severely impaired. Expression of a heterologous c-di-GMP phosphodiesterase restored colonization, demonstrating that the effect is due to high c-di-GMP levels. In the constitutive high-c-di-GMP state, colonizing V. fischeri exhibited reduced motility, altered biofilm aggregate morphology, and a regulatory interaction where transcription of one polysaccharide locus is inhibited by the presence of the other polysaccharide. Our results highlight the importance of proper c-di-GMP regulation during beneficial animal colonization, illustrate multiple pathways regulated by c-di-GMP in the host, and uncover an interplay of multiple exopolysaccharide systems in host-associated aggregates. IMPORTANCE There is substantial interest in studying cyclic diguanylate (c-di-GMP) in pathogenic and environmental bacteria, which has led to an accepted paradigm in which high c-di-GMP levels promote biofilm formation and reduce motility. However, considerably less focus has been placed on understanding how this compound contributes to beneficial colonization. Using the Vibrio fischeri-Hawaiian bobtail squid study system, we took advantage of recent genetic advances in the bacterium to modulate c-di-GMP levels and measure colonization and track c-di-GMP phenotypes in a symbiotic interaction. Studies in the animal host revealed a c-di-GMP-dependent genetic interaction between two distinct biofilm polysaccharides, Syp and cellulose, that was not evident in culture-based studies: elevated c-di-GMP altered the composition and abundance of the in vivo biofilm by decreasing syp transcription due to increased cellulose synthesis. This study reveals important parallels between pathogenic and beneficial colonization and additionally identifies c-di-GMP-dependent regulation that occurs specifically in the squid host.https://journals.asm.org/doi/10.1128/mbio.01671-22Vibrio fischerimicrobiomeEuprymna scolopessquidc-di-GMPbiofilm |
| spellingShingle | Ruth Y. Isenberg David G. Christensen Karen L. Visick Mark J. Mandel High Levels of Cyclic Diguanylate Interfere with Beneficial Bacterial Colonization mBio Vibrio fischeri microbiome Euprymna scolopes squid c-di-GMP biofilm |
| title | High Levels of Cyclic Diguanylate Interfere with Beneficial Bacterial Colonization |
| title_full | High Levels of Cyclic Diguanylate Interfere with Beneficial Bacterial Colonization |
| title_fullStr | High Levels of Cyclic Diguanylate Interfere with Beneficial Bacterial Colonization |
| title_full_unstemmed | High Levels of Cyclic Diguanylate Interfere with Beneficial Bacterial Colonization |
| title_short | High Levels of Cyclic Diguanylate Interfere with Beneficial Bacterial Colonization |
| title_sort | high levels of cyclic diguanylate interfere with beneficial bacterial colonization |
| topic | Vibrio fischeri microbiome Euprymna scolopes squid c-di-GMP biofilm |
| url | https://journals.asm.org/doi/10.1128/mbio.01671-22 |
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