The Glaesserella parasuis phosphoglucomutase is partially required for lipooligosaccharide synthesis
Abstract Lipooligosaccharides (LOSs) are virulence determinants of Glaesserella parasuis, a pathogen of the respiratory tract of pigs. We previously reported that disruption of the galU or galE gene in G. parasuis results in increased sensitivity to porcine serum, indicating that the galactose catab...
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| Format: | Article |
| Language: | English |
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BMC
2020-07-01
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| Series: | Veterinary Research |
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| Online Access: | http://link.springer.com/article/10.1186/s13567-020-00822-9 |
| _version_ | 1828471446923378688 |
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| author | Saixiang Feng Aihua Chen Xiaobing Wang Zhichao Pan Siqi Xu Huiwen Yu Bin Zhang Ming Liao |
| author_facet | Saixiang Feng Aihua Chen Xiaobing Wang Zhichao Pan Siqi Xu Huiwen Yu Bin Zhang Ming Liao |
| author_sort | Saixiang Feng |
| collection | DOAJ |
| description | Abstract Lipooligosaccharides (LOSs) are virulence determinants of Glaesserella parasuis, a pathogen of the respiratory tract of pigs. We previously reported that disruption of the galU or galE gene in G. parasuis results in increased sensitivity to porcine serum, indicating that the galactose catabolism pathway is required for polysaccharide formation in G. parasuis. Here, we evaluated the role of the HAPS_0849 gene in LOS synthesis. The G. parasuis SC096 HAPS_0849 mutant produced a highly truncated LOS molecule, although a small fraction of intact LOS was still observed, and this mutant was found to be more sensitive to serum than the parental strain. HAPS_0849 was overexpressed and purified for biochemical assays, and this protein exhibited phosphoglucomutase (PGM) activity. Heterologous expression of a pgm gene from Escherichia coli in the HAPS_0849 mutant led to restoration of the wild-type LOS glycoform, further demonstrating the PGM function of HAPS_0849 in G. parasuis. The autoagglutination and biofilm formation ability of this strain were also investigated. Disruption of HAPS_0849 led to an increased tendency to autoagglutinate and form more biofilms, and these enhanced phenotypes were observed in the absence of glucose. In addition, LOSs from HAPS_0849, galU and lgtB mutants had similar truncated glycoforms, while LOSs from the galE and lex-1 mutants exhibited another type of defective LOS pattern. These findings imply that HAPS_0849 may function upstream of GalU in the generation of glucose 1-phosphate. In conclusion, our results preliminarily described the functions of HAPS_0849 in G. parasuis, and this gene was partially required for LOS synthesis. |
| first_indexed | 2024-12-11T05:11:27Z |
| format | Article |
| id | doaj.art-d4000c064ac14c298a6ba405af700470 |
| institution | Directory Open Access Journal |
| issn | 1297-9716 |
| language | English |
| last_indexed | 2024-12-11T05:11:27Z |
| publishDate | 2020-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Veterinary Research |
| spelling | doaj.art-d4000c064ac14c298a6ba405af7004702022-12-22T01:19:54ZengBMCVeterinary Research1297-97162020-07-0151111210.1186/s13567-020-00822-9The Glaesserella parasuis phosphoglucomutase is partially required for lipooligosaccharide synthesisSaixiang Feng0Aihua Chen1Xiaobing Wang2Zhichao Pan3Siqi Xu4Huiwen Yu5Bin Zhang6Ming Liao7College of Veterinary Medicine, South China Agricultural UniversityCollege of Veterinary Medicine, South China Agricultural UniversityCollege of Veterinary Medicine, South China Agricultural UniversityCollege of Veterinary Medicine, South China Agricultural UniversityCollege of Veterinary Medicine, South China Agricultural UniversityCollege of Veterinary Medicine, South China Agricultural UniversityCollege of Life Science and Technology, Southwest Minzu UniversityCollege of Veterinary Medicine, South China Agricultural UniversityAbstract Lipooligosaccharides (LOSs) are virulence determinants of Glaesserella parasuis, a pathogen of the respiratory tract of pigs. We previously reported that disruption of the galU or galE gene in G. parasuis results in increased sensitivity to porcine serum, indicating that the galactose catabolism pathway is required for polysaccharide formation in G. parasuis. Here, we evaluated the role of the HAPS_0849 gene in LOS synthesis. The G. parasuis SC096 HAPS_0849 mutant produced a highly truncated LOS molecule, although a small fraction of intact LOS was still observed, and this mutant was found to be more sensitive to serum than the parental strain. HAPS_0849 was overexpressed and purified for biochemical assays, and this protein exhibited phosphoglucomutase (PGM) activity. Heterologous expression of a pgm gene from Escherichia coli in the HAPS_0849 mutant led to restoration of the wild-type LOS glycoform, further demonstrating the PGM function of HAPS_0849 in G. parasuis. The autoagglutination and biofilm formation ability of this strain were also investigated. Disruption of HAPS_0849 led to an increased tendency to autoagglutinate and form more biofilms, and these enhanced phenotypes were observed in the absence of glucose. In addition, LOSs from HAPS_0849, galU and lgtB mutants had similar truncated glycoforms, while LOSs from the galE and lex-1 mutants exhibited another type of defective LOS pattern. These findings imply that HAPS_0849 may function upstream of GalU in the generation of glucose 1-phosphate. In conclusion, our results preliminarily described the functions of HAPS_0849 in G. parasuis, and this gene was partially required for LOS synthesis.http://link.springer.com/article/10.1186/s13567-020-00822-9Glaesserella parasuislipooligosaccharidephosphoglucomutasebiofilm |
| spellingShingle | Saixiang Feng Aihua Chen Xiaobing Wang Zhichao Pan Siqi Xu Huiwen Yu Bin Zhang Ming Liao The Glaesserella parasuis phosphoglucomutase is partially required for lipooligosaccharide synthesis Veterinary Research Glaesserella parasuis lipooligosaccharide phosphoglucomutase biofilm |
| title | The Glaesserella parasuis phosphoglucomutase is partially required for lipooligosaccharide synthesis |
| title_full | The Glaesserella parasuis phosphoglucomutase is partially required for lipooligosaccharide synthesis |
| title_fullStr | The Glaesserella parasuis phosphoglucomutase is partially required for lipooligosaccharide synthesis |
| title_full_unstemmed | The Glaesserella parasuis phosphoglucomutase is partially required for lipooligosaccharide synthesis |
| title_short | The Glaesserella parasuis phosphoglucomutase is partially required for lipooligosaccharide synthesis |
| title_sort | glaesserella parasuis phosphoglucomutase is partially required for lipooligosaccharide synthesis |
| topic | Glaesserella parasuis lipooligosaccharide phosphoglucomutase biofilm |
| url | http://link.springer.com/article/10.1186/s13567-020-00822-9 |
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