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

<p dir="ltr"><i>Helicobacter pylori</i> is a pathogenic bacterium associated with developing gastric cancer and other gastric disorders. One of its major virulence factors, lipopolysaccharide (LPS), plays a crucial role in maintaining bacterial integrity, mediating host a...

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المؤلف الرئيسي: Mou-Chieh Kao (18952576) (author)
منشور في: 2025
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author Mou-Chieh Kao (18952576)
author_facet Mou-Chieh Kao (18952576)
author_role author
dc.creator.none.fl_str_mv Mou-Chieh Kao (18952576)
dc.date.none.fl_str_mv 2025-01-08T03:13:15Z
dc.identifier.none.fl_str_mv 10.6084/m9.figshare.28081937.v1
dc.relation.none.fl_str_mv https://figshare.com/articles/dataset/Roles_of_the_lipopolysaccharide_biosynthesis-related_gene_i_HP0858_i_in_the_fitness_of_i_Helicobacter_pylori_i_and_its_virulence_in_i_Galleria_mellonella_i_/28081937
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Bacteriology
Helicobacter pylori
lipopolysaccharide
ADP-heptose
bi-functional kinase/adenylyltransferase (RfaE/HldE/HP0858)
adhesin AlpA
protein glycosylation
Galleria mellonella
bacterial virulence
drug target
dc.title.none.fl_str_mv Roles of the lipopolysaccharide biosynthesis-related gene <i>HP0858</i> in the fitness of <i>Helicobacter pylori </i>and its virulence in <i>Galleria mellonella</i>
dc.type.none.fl_str_mv Dataset
info:eu-repo/semantics/publishedVersion
dataset
description <p dir="ltr"><i>Helicobacter pylori</i> is a pathogenic bacterium associated with developing 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 LPS synthesis cascade, is a novel pathogen-associated molecular pattern for <i>H. pylori</i>. This study focuses on the <i>HP0858</i> gene, which is predicted to encode RfaE/HldE, an enzyme with kinase and ADP-transferase activities essential for heptose production. An <i>HP0858</i> gene-disrupted mutant was first generated, and the resulting mutant exhibited a truncated LPS structure, confirming its role in LPS biosynthesis. The <i>HP0858</i>-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 a failure to induce IL-8 secretion in infected gastric AGS cells. In an <i>in vivo</i> <i>Galleria mellonella</i> infection model, the <i>HP0858</i> 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 <i>HP0858</i> 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 <i>H. pylori</i> infections.</p>
eu_rights_str_mv openAccess
id Manara_fbbefde324ffc1d2ca8e78fcc8d86cc4
identifier_str_mv 10.6084/m9.figshare.28081937.v1
network_acronym_str Manara
network_name_str ManaraRepo
oai_identifier_str oai:figshare.com:article/28081937
publishDate 2025
repository.mail.fl_str_mv
repository.name.fl_str_mv
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rights_invalid_str_mv CC BY 4.0
spelling Roles of the lipopolysaccharide biosynthesis-related gene <i>HP0858</i> in the fitness of <i>Helicobacter pylori </i>and its virulence in <i>Galleria mellonella</i>Mou-Chieh Kao (18952576)BacteriologyHelicobacter pylorilipopolysaccharideADP-heptosebi-functional kinase/adenylyltransferase (RfaE/HldE/HP0858)adhesin AlpAprotein glycosylationGalleria mellonellabacterial virulencedrug target<p dir="ltr"><i>Helicobacter pylori</i> is a pathogenic bacterium associated with developing 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 LPS synthesis cascade, is a novel pathogen-associated molecular pattern for <i>H. pylori</i>. This study focuses on the <i>HP0858</i> gene, which is predicted to encode RfaE/HldE, an enzyme with kinase and ADP-transferase activities essential for heptose production. An <i>HP0858</i> gene-disrupted mutant was first generated, and the resulting mutant exhibited a truncated LPS structure, confirming its role in LPS biosynthesis. The <i>HP0858</i>-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 a failure to induce IL-8 secretion in infected gastric AGS cells. In an <i>in vivo</i> <i>Galleria mellonella</i> infection model, the <i>HP0858</i> 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 <i>HP0858</i> 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 <i>H. pylori</i> infections.</p>2025-01-08T03:13:15ZDatasetinfo:eu-repo/semantics/publishedVersiondataset10.6084/m9.figshare.28081937.v1https://figshare.com/articles/dataset/Roles_of_the_lipopolysaccharide_biosynthesis-related_gene_i_HP0858_i_in_the_fitness_of_i_Helicobacter_pylori_i_and_its_virulence_in_i_Galleria_mellonella_i_/28081937CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/280819372025-01-08T03:13:15Z
spellingShingle Roles of the lipopolysaccharide biosynthesis-related gene <i>HP0858</i> in the fitness of <i>Helicobacter pylori </i>and its virulence in <i>Galleria mellonella</i>
Mou-Chieh Kao (18952576)
Bacteriology
Helicobacter pylori
lipopolysaccharide
ADP-heptose
bi-functional kinase/adenylyltransferase (RfaE/HldE/HP0858)
adhesin AlpA
protein glycosylation
Galleria mellonella
bacterial virulence
drug target
status_str publishedVersion
title Roles of the lipopolysaccharide biosynthesis-related gene <i>HP0858</i> in the fitness of <i>Helicobacter pylori </i>and its virulence in <i>Galleria mellonella</i>
title_full Roles of the lipopolysaccharide biosynthesis-related gene <i>HP0858</i> in the fitness of <i>Helicobacter pylori </i>and its virulence in <i>Galleria mellonella</i>
title_fullStr Roles of the lipopolysaccharide biosynthesis-related gene <i>HP0858</i> in the fitness of <i>Helicobacter pylori </i>and its virulence in <i>Galleria mellonella</i>
title_full_unstemmed Roles of the lipopolysaccharide biosynthesis-related gene <i>HP0858</i> in the fitness of <i>Helicobacter pylori </i>and its virulence in <i>Galleria mellonella</i>
title_short Roles of the lipopolysaccharide biosynthesis-related gene <i>HP0858</i> in the fitness of <i>Helicobacter pylori </i>and its virulence in <i>Galleria mellonella</i>
title_sort Roles of the lipopolysaccharide biosynthesis-related gene <i>HP0858</i> in the fitness of <i>Helicobacter pylori </i>and its virulence in <i>Galleria mellonella</i>
topic Bacteriology
Helicobacter pylori
lipopolysaccharide
ADP-heptose
bi-functional kinase/adenylyltransferase (RfaE/HldE/HP0858)
adhesin AlpA
protein glycosylation
Galleria mellonella
bacterial virulence
drug target