Glycosyltransferase Jhp0106 (PseE) contributes to flagellin maturation in Helicobacter pylori

Autor:	Pei Jane Tsai, Yueh Lin Chen, Cheng Yen Kao, Marcia Shu-Wei Su, Shuying Wang, Ching Hao Teng, Shiau Ting Hu, Jiunn Jong Wu, Kai Yuan Yang, Jenn Wei Chen
Jazyk:	angličtina
Rok vydání:	2021
Předmět:	Glycosylation Mutant Virulence Motility Flagellum flagellin glycosyltransferase Helicobacter Infections 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Glycosyltransferase Humans Amino Acid Sequence biology Helicobacter pylori Chemistry Gastroenterology Active site Jhp0106 Glycosyltransferases General Medicine Original Articles bacterial infections and mycoses Infectious Diseases Biochemistry motility Flagella 030220 oncology & carcinogenesis biology.protein bacteria 030211 gastroenterology & hepatology Original Article Flagellin
Zdroj:	Helicobacter
ISSN:	1523-5378 1083-4389
Popis:	Background Flagella‐mediated motility is both a crucial virulence determinant of Helicobacter pylori and a factor associated with gastrointestinal diseases. Flagellar formation requires flagellins to be glycosylated with pseudaminic acid (Pse), a process that has been extensively studied. However, the transfer of Pse to flagellins remains poorly understood. Therefore, the aim of this study is to characterize a putative glycosyltransferase jhp0106 in flagellar formation. Materials and Methods Western blotting and chemical deglycosylation were performed to examine FlaA glycosylation. Protein structural analyses were executed to identify the active site residues of Jhp0106, while the Jhp0106‐FlaA interaction was examined using a bacterial two‐hybrid assay. Lastly, site‐directed mutants with mutated active site residues in the jhp0106 gene were generated and investigated using a motility assay, Western blotting, cDNA‐qPCR analysis, and electron microscopic examination. Results Loss of flagellar formation in the Δjhp0106 mutant was confirmed to be associated with non‐glycosylated FlaA. Furthermore, three active site residues of Jhp0106 (S350, F376, and E415) were identified within a potential substrate‐binding region. The interaction between FlaA and Jhp0106, Jhp0106::S350A, Jhp0106::F376A, or Jhp0106::E415A was determined to be significant. As well, the substitution of S350A, F376A, or E415A in the site‐directed Δjhp0106 mutants resulted in impaired motility, deficient FlaA glycosylation, and lacking flagella. However, these phenotypic changes were regardless of flaA expression, implying an indefinite proteolytic degradation of FlaA occurred. Conclusions This study demonstrated that Jhp0106 (PseE) binds to FlaA mediating FlaA glycosylation and flagellar formation. Our discovery of PseE has revealed a new glycosyltransferase family responsible for flagellin glycosylation in pathogens.
Databáze:	OpenAIRE
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