Downregulated Expression of Virulence Factors Induced by Benzyl Isothiocyanate in Staphylococcus Aureus: A Transcriptomic Analysis

Int J Mol Sci. 2019 Oct 31;20(21):5441. doi: 10.3390/ijms20215441.

Abstract

Staphylococcus aureus (S. aureus) is a common foodborne pathogen that leads to various diseases; therefore, we urgently need to identify different means to control this harmful pathogen in food. In this study, we monitored the transcriptional changes of S. aureus by RNA-seq analysis to better understand the effect of benzyl isothiocyanate (BITC) on the virulence inhibition of S. aureus and determined the bacteriostatic effect of BITC at subinhibitory concentrations. Our results revealed that, compared with the control group (SAC), the BITC-treated experimental group (SAQ_BITC) had 708 differentially expressed genes (DEGs), of which 333 genes were downregulated and the capsular polysaccharide (cp) was significantly downregulated. Furthermore, we screened five of the most virulent factors of S. aureus, including the capsular polysaccharide biosynthesis protein (cp5D), capsular polysaccharide synthesis enzyme (cp8F), thermonuclease (nuc), clumping factor (clf), and protein A (spa), and verified the accuracy of these significantly downregulated genes by qRT-PCR. At the same time, we used light microscopy, scanning electron microscopy (SEM) and inverted fluorescence microscopy (IFM) to observe changes in biofilm associated with the cp5D and cp8F. Therefore, these results will help to further study the basis of BITC for the antibacterial action of foodborne pathogenic bacteria.

Keywords: RNA-seq; Staphylococcus aureus; benzyl isothiocyanate; biofilm; differentially expressed genes.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Biofilms / drug effects
  • Biofilms / growth & development
  • Down-Regulation / drug effects
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Bacterial / drug effects*
  • Isothiocyanates / pharmacology*
  • Microbial Sensitivity Tests
  • Microscopy, Electron, Scanning
  • Microscopy, Fluorescence
  • Sequence Analysis, RNA / methods
  • Staphylococcus aureus / genetics*
  • Staphylococcus aureus / pathogenicity
  • Staphylococcus aureus / physiology
  • Virulence / genetics
  • Virulence Factors / genetics

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Isothiocyanates
  • Virulence Factors
  • benzyl isothiocyanate