DOI: 10.5061/dryad.2bvq83c38
title: Data from: Mucus-derived glycans are inhibitory signals for <em>Salmonella</em> Typhimurium SPI-1-mediated invasion

description: Mucus forms a critical barrier against enteric pathogens like Salmonella
 enterica serovar Typhimurium. While in vivo studies indicate that
 secreted, gel-forming mucins and specifically Core 3 glycosylation are
 protective against S. Typhimurium, the molecular mechanisms involved
 remain unclear. Here, we demonstrate that native intestinal mucins inhibit
 Salmonella invasion of colonic epithelial cells by downregulating the type
 3 secretion system (T3SS) through suppression of the key virulence
 regulator, HilD. By measuring changes in the transcriptional profile
 (qPCR, RNA-seq), proteomics (DIA-based), growth, and HT-29 host cell
 invasion, our study identifies mucin glycans and specific mucin sugars,
 namely N-acetyl galactosamine (GalNAc) and N-acetyl glucosamine (GlcNAc),
 as the components responsible for mucin’s anti-virulence effect, likely
 via interaction with HilD's putative carbohydrate-binding domain.
 Notably, we find the native presentation of these sugars is important for
 activity. These insights provide a mechanistic foundation for mucin-based
 strategies to combat enteric infections and, given the prevalence of
 homologous AraC-type regulators in other pathogens, suggest mucins'
 potential as broad-spectrum anti-virulence agents.
description: # Data from: Mucus-derived glycans are inhibitory signals for *Salmonella*
 Typhimurium SPI-1-mediated invasion Dataset DOI:
 [10.5061/dryad.2bvq83c38](https://doi.org/10.5061/dryad.2bvq83c38) ##
 Description of the data and file structure These are Excel files
 containing the numerical data used to generate the final figures in the
 manuscript. Specific experimental efforts represented in this dataset
 include *Salmonella* invasion assays, qPCR for SPI-1 gene expression,
 enumeration of colony-forming units to monitor Salmonella growth, and
 DIA-based proteomics. ### Files and variables ##### File: Figure_1.xlsx
 **Description:** Invasion assays. Log2 fold change in invaded Salmonella
 cells after 1 h treatment with MUC2 (tab: 1B) or MUC5AC (tab: 1C) relative
 to medium alone. Columns, Strain ID. ##### File: Figure_2.xlsx
 **Description:** qPCR data. Log2(fold change in gene expression with MUC2
 relative to medium alone) (tab: 2A). RNA-seq data underlying the volcano
 plot. For each transcript, -log10 False discovery rate, Log2(fold change
 in gene expression) with MUC2 vs. Medium alone (tab: 2B). qPCR data.
 Log2(fold change in gene expression with MUC5AC relative to medium alone)
 (tab: 2C). Growth curve data. The number of colony-forming units at each
 time point with or without MUC2 (tab: 2D). Proteomics data underlying the
 volcano plot. For each protein, -log10 p-value, Log2(fold change in
 expression) with MUC2 vs. Medium alone (tab: 2e). ##### File:
 Figure_3.xlsx **Description:**  Invasion assays. Log2 fold change in
 invaded Salmonella cells after 1 h treatment with 0.05% MUC2 glycans (tab:
 3B) Columns, Strain ID. qPCR data. Log2(fold change in gene expression
 with 0.05% MUC2 glycans or 0.1% Monosaccharides relative to medium alone)
 (tab: 3C). RNA-seq data underlying the volcano plot. For each transcript,
 -log10 False discovery rate, Log2(fold change in gene expression) with
 0.1% MUC2 glycans vs. Medium alone (tab: 3C). ##### File: Figure_4.xlsx
 **Description:** qPCR data. Log2(fold change in hilA expression with MUC2
 relative to medium alone), Columns=Mutant ID (tab: 4B). qPCR data.
 Log2(fold change in gene expression with GalNAc or GlcNAc monosaccharides
 relative to medium alone) in two different strain backgrounds, WT of
 HilD^Q39E,N44D,H95L^ (tab: 4E). ##### File: Figure_5.xlsx
 **Description:** RNA-seq data underlying the heat map. Log2(fold change in
 gene expression relative to medium alone) Rows = gene name, Column =
 treatment (i.e., MUC2, glycans, Monsaccharide pool, individual
 monosaccharides) (tab: 5A).  qPCR data. Log2(fold change in hilA
 expression relative to medium alone), Columns=Treatment type & strain
 background (tab: 5C). qPCR data. Log2(fold change in prgH expression
 relative to medium alone), Columns=Treatment type & strain background
 (tab: 5D). Invasion assays. Number of colony forming units of invaded
 Salmonella cells after 1 h treatment with medium alone or
 glycopolypeptides. Rows = Strain (WT, ∆hilD, or HilD-mutant), Column =
 treatment (tab: 5E) ##### File: Figure_6.xlsx **Description:** qPCR data.
 Log2(fold change in hilA expression relative to medium alone),
 Columns=strain background, Row = treatment type (tab: 6B). qPCR data.
 Log2(fold change in prgH expression relative to medium alone),
 Columns=strain background, Row = treatment type (tab: 6C) ## Access
 information Other publicly accessible locations of the data: * n/a Data
 was derived from the following sources: * n/a