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Related Concept Videos

NF-κB-dependent Signaling Pathway02:26

NF-κB-dependent Signaling Pathway

The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
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Types of RNA

Overview
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Translational Regulation01:29

Translational Regulation

Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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Transcriptional Regulation: Riboswitches

Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...
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NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells
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Bacterial effector binding to ribosomal protein s3 subverts NF-kappaB function.

Xiaofei Gao1, Fengyi Wan, Kristina Mateo

  • 1Department of Microbiology, Molecular Genetics, and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA.

Plos Pathogens
|December 31, 2009
PubMed
Summary

Enterohemorrhagic Escherichia coli (EHEC) virulence factors NleH1 and NleH2 bind to human ribosomal protein S3 (RPS3), impacting host cell transcription and immune responses. Deleting nleH1 increased EHEC virulence in a piglet model.

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Area of Science:

  • Microbiology and Immunology
  • Molecular Biology
  • Pathogen-Host Interactions

Background:

  • Enteric bacterial pathogens, including Enterohemorrhagic Escherichia coli (EHEC), cause significant global health and economic burdens through foodborne illnesses.
  • EHEC utilizes a type III secretion system (T3SS) to inject effector proteins into host cells, subverting cellular functions to promote disease, yet effector mechanisms remain incompletely understood.
  • EHEC infection can lead to severe bloody diarrhea and potentially fatal hemolytic uremic syndrome, with no current effective treatments or prophylaxis.

Purpose of the Study:

  • To investigate the interaction between E. coli O157:H7 T3SS effectors NleH1 and NleH2 and host cell proteins.
  • To elucidate the functional consequences of NleH1 and NleH2 binding to host targets on cellular processes, particularly transcriptional regulation.
  • To assess the role of the nleH1 gene in EHEC virulence using a relevant animal model.

Main Methods:

  • Co-localization studies to determine the cellular location of NleH1, NleH2, and human ribosomal protein S3 (RPS3).
  • Binding assays to identify the regions of NleH1, NleH2, and RPS3 involved in their interaction.
  • Kinase activity assays for NleH1 and NleH2, and reporter gene assays to assess transcriptional regulation dependent on RPS3 and NF-kappaB.
  • In vivo virulence studies in a gnotobiotic piglet model of Shiga toxin-producing E. coli infection with and without the nleH1 gene.

Main Results:

  • E. coli O157:H7 T3SS effectors NleH1 and NleH2 bind to human RPS3, a component of NF-kappaB transcriptional complexes, localizing to the cytoplasm.
  • NleH1 binding to RPS3 reduced nuclear RPS3 abundance and repressed RPS3/NF-kappaB-dependent transcription, while NleH2 stimulated both RPS3-dependent and AP-1-dependent transcription.
  • Deletion of nleH1 from E. coli O157:H7 resulted in a hypervirulent phenotype in a gnotobiotic piglet model, suggesting NleH1 negatively regulates virulence.

Conclusions:

  • EHEC NleH1 and NleH2 are T3SS effectors that modulate host cell transcription by interacting with RPS3, a key component of NF-kappaB signaling.
  • NleH1 acts as a repressor of RPS3/NF-kappaB-dependent transcription, contributing to virulence, while NleH2 has distinct stimulatory effects.
  • These findings suggest that NleH effectors disrupt host innate immune responses by interfering with transcriptional complexes, highlighting a novel virulence mechanism for EHEC.