Characterizing the Effect of the Lysine Deacetylation Modification on Enzyme Activity of Pyruvate Kinase I and Pathogenicity of Vibrio alginolyticus
Pyruvate kinase I (PykF) is one of the key enzymes of glycolysis and plays a crucial role in bacterial metabolism. Several acetylation sites of Vibrio alginolyticus PykF were reported in previous studies and then 11 sites were first verified in this study, however, the specific roles of PykF acetyla...
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Frontiers Media S.A.
2022-06-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2022.877067/full |
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| author | Zhou Xu Zhou Xu Zhou Xu Linjing Wang Linjing Wang Linjing Wang Xudong Wang Xudong Wang Xudong Wang Mingyue Wan Mingyue Wan Mingyue Wan Mei Tang Mei Tang Mei Tang Yu Ding Yu Ding Yu Ding |
| author_facet | Zhou Xu Zhou Xu Zhou Xu Linjing Wang Linjing Wang Linjing Wang Xudong Wang Xudong Wang Xudong Wang Mingyue Wan Mingyue Wan Mingyue Wan Mei Tang Mei Tang Mei Tang Yu Ding Yu Ding Yu Ding |
| author_sort | Zhou Xu |
| collection | DOAJ |
| description | Pyruvate kinase I (PykF) is one of the key enzymes of glycolysis and plays a crucial role in bacterial metabolism. Several acetylation sites of Vibrio alginolyticus PykF were reported in previous studies and then 11 sites were first verified in this study, however, the specific roles of PykF acetylation remains unclear. Overlap-PCR and homologous recombination were implied to delete V. alginolyticus pykF gene and constructed complementary strains of site-directed mutagenesis for the further research focus on the deacetylation regulation on PykF. The results showed that the pyruvate kinase activity was sharply suppressed in the deacetylation status of K52, K68, and K317 of PykF, as well as the extracellular protease activity was significantly decreased in the deacetylation status of K52 and K68, but not induced with K317. Moreover, the growth rates of V. alginolyticus were not influenced with these three deacetylation sites. The ΔpykF mutant exhibited a 6-fold reduction in virulence to zebrafish. Site-directed mutations of K52R and K68R also showed reduced virulence while mutations of K317R didn't. The in vitro experiments showed that PykF was acetylated by acetyl phosphate (AcP), with the increase of incubation time by AcP, the acetylation level of PykF increased while the enzyme activity of PykF decreased correspondingly. Besides, PykF was deacetylated by CobB deacetylase and in result that the deacetylation was significantly down-regulated while the pyruvate kinase activity of PykF increased. Moreover, deletion of cobB gene had no significant difference in pyruvate kinase activity. These results confirm that CobB can regulate the acetylation level and pyruvate kinase activity of PykF. In summary, the results of this study provide a theoretical basis for further understanding of the deacetylation modification of PykF. It provides a new idea for the prevention and cure of vibriosis. |
| first_indexed | 2024-12-12T03:47:48Z |
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| id | doaj.art-52cb7bb5918441f69cdbebcf745034b2 |
| institution | Directory Open Access Journal |
| issn | 2297-1769 |
| language | English |
| last_indexed | 2024-12-12T03:47:48Z |
| publishDate | 2022-06-01 |
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| series | Frontiers in Veterinary Science |
| spelling | doaj.art-52cb7bb5918441f69cdbebcf745034b22022-12-22T00:39:28ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692022-06-01910.3389/fvets.2022.877067877067Characterizing the Effect of the Lysine Deacetylation Modification on Enzyme Activity of Pyruvate Kinase I and Pathogenicity of Vibrio alginolyticusZhou Xu0Zhou Xu1Zhou Xu2Linjing Wang3Linjing Wang4Linjing Wang5Xudong Wang6Xudong Wang7Xudong Wang8Mingyue Wan9Mingyue Wan10Mingyue Wan11Mei Tang12Mei Tang13Mei Tang14Yu Ding15Yu Ding16Yu Ding17Fisheries College, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, ChinaGuangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang, ChinaFisheries College, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, ChinaGuangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang, ChinaFisheries College, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, ChinaGuangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang, ChinaFisheries College, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, ChinaGuangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang, ChinaFisheries College, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, ChinaGuangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang, ChinaFisheries College, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, ChinaGuangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang, ChinaPyruvate kinase I (PykF) is one of the key enzymes of glycolysis and plays a crucial role in bacterial metabolism. Several acetylation sites of Vibrio alginolyticus PykF were reported in previous studies and then 11 sites were first verified in this study, however, the specific roles of PykF acetylation remains unclear. Overlap-PCR and homologous recombination were implied to delete V. alginolyticus pykF gene and constructed complementary strains of site-directed mutagenesis for the further research focus on the deacetylation regulation on PykF. The results showed that the pyruvate kinase activity was sharply suppressed in the deacetylation status of K52, K68, and K317 of PykF, as well as the extracellular protease activity was significantly decreased in the deacetylation status of K52 and K68, but not induced with K317. Moreover, the growth rates of V. alginolyticus were not influenced with these three deacetylation sites. The ΔpykF mutant exhibited a 6-fold reduction in virulence to zebrafish. Site-directed mutations of K52R and K68R also showed reduced virulence while mutations of K317R didn't. The in vitro experiments showed that PykF was acetylated by acetyl phosphate (AcP), with the increase of incubation time by AcP, the acetylation level of PykF increased while the enzyme activity of PykF decreased correspondingly. Besides, PykF was deacetylated by CobB deacetylase and in result that the deacetylation was significantly down-regulated while the pyruvate kinase activity of PykF increased. Moreover, deletion of cobB gene had no significant difference in pyruvate kinase activity. These results confirm that CobB can regulate the acetylation level and pyruvate kinase activity of PykF. In summary, the results of this study provide a theoretical basis for further understanding of the deacetylation modification of PykF. It provides a new idea for the prevention and cure of vibriosis.https://www.frontiersin.org/articles/10.3389/fvets.2022.877067/fullVibrio alginolyticuslysine deacetylationPykFpost-translational modificationglycolysis |
| spellingShingle | Zhou Xu Zhou Xu Zhou Xu Linjing Wang Linjing Wang Linjing Wang Xudong Wang Xudong Wang Xudong Wang Mingyue Wan Mingyue Wan Mingyue Wan Mei Tang Mei Tang Mei Tang Yu Ding Yu Ding Yu Ding Characterizing the Effect of the Lysine Deacetylation Modification on Enzyme Activity of Pyruvate Kinase I and Pathogenicity of Vibrio alginolyticus Frontiers in Veterinary Science Vibrio alginolyticus lysine deacetylation PykF post-translational modification glycolysis |
| title | Characterizing the Effect of the Lysine Deacetylation Modification on Enzyme Activity of Pyruvate Kinase I and Pathogenicity of Vibrio alginolyticus |
| title_full | Characterizing the Effect of the Lysine Deacetylation Modification on Enzyme Activity of Pyruvate Kinase I and Pathogenicity of Vibrio alginolyticus |
| title_fullStr | Characterizing the Effect of the Lysine Deacetylation Modification on Enzyme Activity of Pyruvate Kinase I and Pathogenicity of Vibrio alginolyticus |
| title_full_unstemmed | Characterizing the Effect of the Lysine Deacetylation Modification on Enzyme Activity of Pyruvate Kinase I and Pathogenicity of Vibrio alginolyticus |
| title_short | Characterizing the Effect of the Lysine Deacetylation Modification on Enzyme Activity of Pyruvate Kinase I and Pathogenicity of Vibrio alginolyticus |
| title_sort | characterizing the effect of the lysine deacetylation modification on enzyme activity of pyruvate kinase i and pathogenicity of vibrio alginolyticus |
| topic | Vibrio alginolyticus lysine deacetylation PykF post-translational modification glycolysis |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2022.877067/full |
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