Bioinformatic and functional characterization of cyclic-di-GMP metabolic proteins in Vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm-associated phenotypes
The biofilm lifestyle is critical for bacterial survival and proliferation in the fluctuating marine environment. Cyclic diguanylate (c-di-GMP) is a key second messenger during bacterial adaptation to various environmental signals, which has been identified as a master regulator of biofilm formation...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-09-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1258415/full |
| _version_ | 1797678007594254336 |
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| author | Xiao-Xiao Gong Xiao-Xiao Gong Xiao-Xiao Gong Yan-Hua Zeng Hai-Min Chen Hai-Min Chen Hai-Min Chen Na Zhang Na Zhang Na Zhang Yue Han Yue Han Yue Han Hao Long Zhen-Yu Xie Zhen-Yu Xie Zhen-Yu Xie |
| author_facet | Xiao-Xiao Gong Xiao-Xiao Gong Xiao-Xiao Gong Yan-Hua Zeng Hai-Min Chen Hai-Min Chen Hai-Min Chen Na Zhang Na Zhang Na Zhang Yue Han Yue Han Yue Han Hao Long Zhen-Yu Xie Zhen-Yu Xie Zhen-Yu Xie |
| author_sort | Xiao-Xiao Gong |
| collection | DOAJ |
| description | The biofilm lifestyle is critical for bacterial survival and proliferation in the fluctuating marine environment. Cyclic diguanylate (c-di-GMP) is a key second messenger during bacterial adaptation to various environmental signals, which has been identified as a master regulator of biofilm formation. However, little is known about whether and how c-di-GMP signaling regulates biofilm formation in Vibrio alginolyticus, a globally dominant marine pathogen. Here, a large set of 63 proteins were predicted to participate in c-di-GMP metabolism (biosynthesis or degradation) in a pathogenic V. alginolyticus strain HN08155. Guided by protein homology, conserved domains and gene context information, a representative subset of 22 c-di-GMP metabolic proteins were selected to determine which ones affect biofilm-associated phenotypes. By comparing phenotypic differences between the wild-type and mutants or overexpression strains, we found that 22 c-di-GMP metabolic proteins can separately regulate different phenotypic outputs in V. alginolyticus. The results indicated that overexpression of four c-di-GMP metabolic proteins, including VA0356, VA1591 (CdgM), VA4033 (DgcB) and VA0088, strongly enhanced rugose colony morphotypes and strengthened Congo Red (CR) binding capacity, both of which are indicators of biofilm matrix overproduction. Furthermore, rugose enhanced colonies were accompanied by increased transcript levels of extracellular polysaccharide (EPS) biosynthesis genes and decreased expression of flagellar synthesis genes compared to smooth colonies (WTpBAD control), as demonstrated by overexpression strains WTp4033 and ∆VA4033p4033. Overall, the high abundance of c-di-GMP metabolic proteins in V. alginolyticus suggests that c-di-GMP signaling and regulatory system could play a key role in its response and adaptation to the ever-changing marine environment. This work provides a robust foundation for the study of the molecular mechanisms of c-di-GMP in the biofilm formation of V. alginolyticus. |
| first_indexed | 2024-03-11T22:53:21Z |
| format | Article |
| id | doaj.art-2559faf2a1364e6d92f50b874aa2b94f |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-03-11T22:53:21Z |
| publishDate | 2023-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-2559faf2a1364e6d92f50b874aa2b94f2023-09-21T22:18:25ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2023-09-011410.3389/fmicb.2023.12584151258415Bioinformatic and functional characterization of cyclic-di-GMP metabolic proteins in Vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm-associated phenotypesXiao-Xiao Gong0Xiao-Xiao Gong1Xiao-Xiao Gong2Yan-Hua Zeng3Hai-Min Chen4Hai-Min Chen5Hai-Min Chen6Na Zhang7Na Zhang8Na Zhang9Yue Han10Yue Han11Yue Han12Hao Long13Zhen-Yu Xie14Zhen-Yu Xie15Zhen-Yu Xie16State Key Laboratory of Marine Resource Utilization in the South China Sea, Hainan University, Haikou, Hainan, ChinaHainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, Hainan University, Haikou, Hainan, ChinaCollege of Marine Sciences, Hainan University, Haikou, Hainan, ChinaState Key Laboratory of Marine Resource Utilization in the South China Sea, Hainan University, Haikou, Hainan, ChinaState Key Laboratory of Marine Resource Utilization in the South China Sea, Hainan University, Haikou, Hainan, ChinaHainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, Hainan University, Haikou, Hainan, ChinaCollege of Marine Sciences, Hainan University, Haikou, Hainan, ChinaState Key Laboratory of Marine Resource Utilization in the South China Sea, Hainan University, Haikou, Hainan, ChinaHainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, Hainan University, Haikou, Hainan, ChinaCollege of Marine Sciences, Hainan University, Haikou, Hainan, ChinaState Key Laboratory of Marine Resource Utilization in the South China Sea, Hainan University, Haikou, Hainan, ChinaHainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, Hainan University, Haikou, Hainan, ChinaCollege of Marine Sciences, Hainan University, Haikou, Hainan, ChinaState Key Laboratory of Marine Resource Utilization in the South China Sea, Hainan University, Haikou, Hainan, ChinaState Key Laboratory of Marine Resource Utilization in the South China Sea, Hainan University, Haikou, Hainan, ChinaHainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, Hainan University, Haikou, Hainan, ChinaCollege of Marine Sciences, Hainan University, Haikou, Hainan, ChinaThe biofilm lifestyle is critical for bacterial survival and proliferation in the fluctuating marine environment. Cyclic diguanylate (c-di-GMP) is a key second messenger during bacterial adaptation to various environmental signals, which has been identified as a master regulator of biofilm formation. However, little is known about whether and how c-di-GMP signaling regulates biofilm formation in Vibrio alginolyticus, a globally dominant marine pathogen. Here, a large set of 63 proteins were predicted to participate in c-di-GMP metabolism (biosynthesis or degradation) in a pathogenic V. alginolyticus strain HN08155. Guided by protein homology, conserved domains and gene context information, a representative subset of 22 c-di-GMP metabolic proteins were selected to determine which ones affect biofilm-associated phenotypes. By comparing phenotypic differences between the wild-type and mutants or overexpression strains, we found that 22 c-di-GMP metabolic proteins can separately regulate different phenotypic outputs in V. alginolyticus. The results indicated that overexpression of four c-di-GMP metabolic proteins, including VA0356, VA1591 (CdgM), VA4033 (DgcB) and VA0088, strongly enhanced rugose colony morphotypes and strengthened Congo Red (CR) binding capacity, both of which are indicators of biofilm matrix overproduction. Furthermore, rugose enhanced colonies were accompanied by increased transcript levels of extracellular polysaccharide (EPS) biosynthesis genes and decreased expression of flagellar synthesis genes compared to smooth colonies (WTpBAD control), as demonstrated by overexpression strains WTp4033 and ∆VA4033p4033. Overall, the high abundance of c-di-GMP metabolic proteins in V. alginolyticus suggests that c-di-GMP signaling and regulatory system could play a key role in its response and adaptation to the ever-changing marine environment. This work provides a robust foundation for the study of the molecular mechanisms of c-di-GMP in the biofilm formation of V. alginolyticus.https://www.frontiersin.org/articles/10.3389/fmicb.2023.1258415/fullVibrio alginolyticusc-di-GMPrugosebiofilmextracellular polysaccharide |
| spellingShingle | Xiao-Xiao Gong Xiao-Xiao Gong Xiao-Xiao Gong Yan-Hua Zeng Hai-Min Chen Hai-Min Chen Hai-Min Chen Na Zhang Na Zhang Na Zhang Yue Han Yue Han Yue Han Hao Long Zhen-Yu Xie Zhen-Yu Xie Zhen-Yu Xie Bioinformatic and functional characterization of cyclic-di-GMP metabolic proteins in Vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm-associated phenotypes Frontiers in Microbiology Vibrio alginolyticus c-di-GMP rugose biofilm extracellular polysaccharide |
| title | Bioinformatic and functional characterization of cyclic-di-GMP metabolic proteins in Vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm-associated phenotypes |
| title_full | Bioinformatic and functional characterization of cyclic-di-GMP metabolic proteins in Vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm-associated phenotypes |
| title_fullStr | Bioinformatic and functional characterization of cyclic-di-GMP metabolic proteins in Vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm-associated phenotypes |
| title_full_unstemmed | Bioinformatic and functional characterization of cyclic-di-GMP metabolic proteins in Vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm-associated phenotypes |
| title_short | Bioinformatic and functional characterization of cyclic-di-GMP metabolic proteins in Vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm-associated phenotypes |
| title_sort | bioinformatic and functional characterization of cyclic di gmp metabolic proteins in vibrio alginolyticus unveils key diguanylate cyclases controlling multiple biofilm associated phenotypes |
| topic | Vibrio alginolyticus c-di-GMP rugose biofilm extracellular polysaccharide |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1258415/full |
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