HilE regulates HilD by blocking DNA binding in Salmonella enterica serovar Typhimurium

Jesse R. Grenz, Jessica E.Cott Chubiz, Pariyamon Thaprawat, James M. Slauch

Research output: Contribution to journalArticle

Abstract

The Salmonella type three secretion system (T3SS), encoded in the Salmonella pathogenicity island 1 (SPI1) locus, mediates the invasion of the host intestinal epithelium. SPI1 expression is dependent upon three AraC-like regulators: HilD, HilC, and RtsA. These regulators act in a complex feed-forward loop to activate each other and hilA, which encodes the activator of the T3SS structural genes. HilD has been shown to be the major integration point of most signals known to activate the expression of the SPI1 T3SS, acting as a switch to control induction of the system. HilE is a negative regulator that acts upon HilD. Here we provide genetic and biochemical data showing that HilE specifically binds to HilD but not to HilC or RtsA. This protein-protein interaction blocks the ability of HilD to bind DNA as shown by both an in vivo reporter system and an in vitro gel shift assay. HilE does not affect HilD dimerization, nor does it control the stability of the HilD protein. We also investigated the role of HilE during the infection of mice using competition assays. Although deletion of hilE does not confer a phenotype, the hilE mutation does suppress the invasion defect conferred by loss of FliZ, which acts as a positive signal controlling HilD protein activity. Together, these data suggest that HilE functions to restrict low-level HilD activity, preventing premature activation of SPI1 until positive inputs reach a threshold required to fully induce the system.

Original languageEnglish (US)
Article numbere00750-17
JournalJournal of bacteriology
Volume200
Issue number8
DOIs
StatePublished - Apr 1 2018

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Salmonella enterica
Genomic Islands
Salmonella
DNA
Proteins
Protein Stability
Dimerization
Intestinal Mucosa
Molecular Biology
Gels
Serogroup
Phenotype
Mutation
Infection
Genes

Keywords

  • HilD
  • HilE
  • SPI1
  • Salmonella

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

HilE regulates HilD by blocking DNA binding in Salmonella enterica serovar Typhimurium. / Grenz, Jesse R.; Chubiz, Jessica E.Cott; Thaprawat, Pariyamon; Slauch, James M.

In: Journal of bacteriology, Vol. 200, No. 8, e00750-17, 01.04.2018.

Research output: Contribution to journalArticle

Grenz, Jesse R. ; Chubiz, Jessica E.Cott ; Thaprawat, Pariyamon ; Slauch, James M. / HilE regulates HilD by blocking DNA binding in Salmonella enterica serovar Typhimurium. In: Journal of bacteriology. 2018 ; Vol. 200, No. 8.
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