A Novel Small RNA Regulates Tolerance and Virulence in Shigella flexneri by Responding to Acidic Environmental Changes

Front Cell Infect Microbiol. 2016 Mar 8:6:24. doi: 10.3389/fcimb.2016.00024. eCollection 2016.

Abstract

Shigella flexneri is an important cause of bacillary dysentery in developing countries. Small regulatory RNAs (sRNAs) play essential roles in diverse cellular processes. We found a novel sRNA Ssr1 based on RT-PCR, northern blot, and 5'RACE in S. flexneri. Ssr1 responds to acidic environmental changes, as shown by a strong linear correlation between the pH value and Ssr1 expression (R = 0.785, P < 0.05) using the qRT-PCR method. Deletion of Ssr1 results in growth retardation at pH values ranging from 5.0 to 7.0 (P < 0.05), and the survival rate was reduced by 22% in acidic conditions (pH 3.0). Additionally, virulence was significantly increased in an Ssr1 mutant strain, as revealed in a murine lung invasion model and survival model assays. By using the sTarPicker method and proteomic analysis, we considered that DnaK, which is a major factor that confers acidic stress tolerance, may be a direct target of Ssr1. We also found that Ssr1 may enhance virulence by directly targeting OmpA; this leads to altered expression of genes in the type three secretion system (T3SS). This work provides new insight into the mechanism of adaptation to environmental stress and into the pathogenesis of Shigella.

Keywords: Shigella flexneri; response; sRNA; stress tolerance; virulence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptation, Physiological / genetics*
  • Animals
  • Bacterial Outer Membrane Proteins / metabolism
  • Dysentery, Bacillary / microbiology
  • Female
  • Gene Expression Regulation, Bacterial / genetics*
  • Hydrogen-Ion Concentration
  • Mice
  • Mice, Inbred BALB C
  • RNA, Bacterial / genetics
  • RNA, Small Untranslated / genetics*
  • Shigella flexneri / genetics*
  • Shigella flexneri / immunology
  • Shigella flexneri / pathogenicity*
  • Transcription, Genetic / genetics
  • Type III Secretion Systems / metabolism
  • Virulence Factors / genetics

Substances

  • Bacterial Outer Membrane Proteins
  • RNA, Bacterial
  • RNA, Small Untranslated
  • Type III Secretion Systems
  • Virulence Factors
  • OMPA outer membrane proteins