Roles of the lipopolysaccharide biosynthesis-related gene HP0858 in the fitness of Helicobacter pylori and its virulence in Galleria mellonella

Helicobacter pylori is a pathogenic bacterium associated with the development of gastric cancer and other gastric disorders. One of its major virulence factors, lipopolysaccharide (LPS), plays a crucial role in maintaining bacterial integrity, mediating host adhesion, and modulating the immune respo...

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Main Authors: Tram Thi Hong Nguyen, Pei-Chun Chen, Po-Chuan Wang, Wen-Han Wang, Chung-Yu Lan, Steffen Backert, Mou-Chieh Kao
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:Virulence
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Online Access:https://www.tandfonline.com/doi/10.1080/21505594.2025.2548620
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author Tram Thi Hong Nguyen
Pei-Chun Chen
Po-Chuan Wang
Wen-Han Wang
Chung-Yu Lan
Steffen Backert
Mou-Chieh Kao
author_facet Tram Thi Hong Nguyen
Pei-Chun Chen
Po-Chuan Wang
Wen-Han Wang
Chung-Yu Lan
Steffen Backert
Mou-Chieh Kao
author_sort Tram Thi Hong Nguyen
collection DOAJ
description Helicobacter pylori is a pathogenic bacterium associated with the development of gastric cancer and other gastric disorders. One of its major virulence factors, lipopolysaccharide (LPS), plays a crucial role in maintaining bacterial integrity, mediating host adhesion, and modulating the immune response. Recent studies have indicated that ADP-heptose, an intermediate in the heptose biosynthetic pathway involved in the LPS synthesis cascade, is a novel pathogen-associated molecular pattern for H. pylori. This study focuses on the HP0858 gene, which is predicted to encode RfaE/HldE, an enzyme with kinase and ADP-transferase activities essential for heptose production. An HP0858 gene-disrupted mutant was first generated, and the resulting mutant exhibited a truncated LPS structure, confirming its role in LPS biosynthesis. The HP0858-deficient mutant displayed increased sensitivity to the detergent SDS and the antibiotic novobiocin, heightened surface hydrophobicity, and a propensity for autoaggregation. Additionally, the mutant exhibited reduced adhesion and internalization capabilities, a diminished elongation phenotype, and failed to induce IL-8 secretion in infected gastric AGS cells. In an in vivo Galleria mellonella infection model, the HP0858 knockout mutant showed significantly attenuated virulence, as no bacterial load was detectable in the larvae’s hemolymph 48 h post-infection, unlike the wild-type strain. Finally, we provided evidence that the enzyme encoded by HP0858 is involved in a general protein glycosylation system linked to LPS biosynthesis, specifically glycosylating the adhesin AlpA. These findings highlight the essential role of RfaE/HldE/HP0858 in LPS biosynthesis and bacterial virulence, making it a promising target for future therapeutic interventions against H. pylori infections.
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spelling doaj-art-4c07c72bbe4f40ec9b32b60ee12d77842025-08-24T16:31:22ZengTaylor & Francis GroupVirulence2150-55942150-56082025-12-0116110.1080/21505594.2025.2548620Roles of the lipopolysaccharide biosynthesis-related gene HP0858 in the fitness of Helicobacter pylori and its virulence in Galleria mellonellaTram Thi Hong Nguyen0Pei-Chun Chen1Po-Chuan Wang2Wen-Han Wang3Chung-Yu Lan4Steffen Backert5Mou-Chieh Kao6Institute of Molecular Medicine, National Tsing Hua University, Hsinchu, TaiwanInstitute of Molecular Medicine, National Tsing Hua University, Hsinchu, TaiwanDepartment of Gastroenterology, Hsinchu MacKay Memorial Hospital, Hsinchu, TaiwanInstitute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu, TaiwanInstitute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu, TaiwanDivision of Microbiology, Department of Biology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyInstitute of Molecular Medicine, National Tsing Hua University, Hsinchu, TaiwanHelicobacter pylori is a pathogenic bacterium associated with the development of gastric cancer and other gastric disorders. One of its major virulence factors, lipopolysaccharide (LPS), plays a crucial role in maintaining bacterial integrity, mediating host adhesion, and modulating the immune response. Recent studies have indicated that ADP-heptose, an intermediate in the heptose biosynthetic pathway involved in the LPS synthesis cascade, is a novel pathogen-associated molecular pattern for H. pylori. This study focuses on the HP0858 gene, which is predicted to encode RfaE/HldE, an enzyme with kinase and ADP-transferase activities essential for heptose production. An HP0858 gene-disrupted mutant was first generated, and the resulting mutant exhibited a truncated LPS structure, confirming its role in LPS biosynthesis. The HP0858-deficient mutant displayed increased sensitivity to the detergent SDS and the antibiotic novobiocin, heightened surface hydrophobicity, and a propensity for autoaggregation. Additionally, the mutant exhibited reduced adhesion and internalization capabilities, a diminished elongation phenotype, and failed to induce IL-8 secretion in infected gastric AGS cells. In an in vivo Galleria mellonella infection model, the HP0858 knockout mutant showed significantly attenuated virulence, as no bacterial load was detectable in the larvae’s hemolymph 48 h post-infection, unlike the wild-type strain. Finally, we provided evidence that the enzyme encoded by HP0858 is involved in a general protein glycosylation system linked to LPS biosynthesis, specifically glycosylating the adhesin AlpA. These findings highlight the essential role of RfaE/HldE/HP0858 in LPS biosynthesis and bacterial virulence, making it a promising target for future therapeutic interventions against H. pylori infections.https://www.tandfonline.com/doi/10.1080/21505594.2025.2548620Helicobacter pylorilipopolysaccharideADP-heptosebi-functional kinase/adenylyltransferase (RfaE/hlde/hp0858)protein glycosylationGalleria mellonella
spellingShingle Tram Thi Hong Nguyen
Pei-Chun Chen
Po-Chuan Wang
Wen-Han Wang
Chung-Yu Lan
Steffen Backert
Mou-Chieh Kao
Roles of the lipopolysaccharide biosynthesis-related gene HP0858 in the fitness of Helicobacter pylori and its virulence in Galleria mellonella
Virulence
Helicobacter pylori
lipopolysaccharide
ADP-heptose
bi-functional kinase/adenylyltransferase (RfaE/hlde/hp0858)
protein glycosylation
Galleria mellonella
title Roles of the lipopolysaccharide biosynthesis-related gene HP0858 in the fitness of Helicobacter pylori and its virulence in Galleria mellonella
title_full Roles of the lipopolysaccharide biosynthesis-related gene HP0858 in the fitness of Helicobacter pylori and its virulence in Galleria mellonella
title_fullStr Roles of the lipopolysaccharide biosynthesis-related gene HP0858 in the fitness of Helicobacter pylori and its virulence in Galleria mellonella
title_full_unstemmed Roles of the lipopolysaccharide biosynthesis-related gene HP0858 in the fitness of Helicobacter pylori and its virulence in Galleria mellonella
title_short Roles of the lipopolysaccharide biosynthesis-related gene HP0858 in the fitness of Helicobacter pylori and its virulence in Galleria mellonella
title_sort roles of the lipopolysaccharide biosynthesis related gene hp0858 in the fitness of helicobacter pylori and its virulence in galleria mellonella
topic Helicobacter pylori
lipopolysaccharide
ADP-heptose
bi-functional kinase/adenylyltransferase (RfaE/hlde/hp0858)
protein glycosylation
Galleria mellonella
url https://www.tandfonline.com/doi/10.1080/21505594.2025.2548620
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