Transfer activation of SXT/R391 integrative and conjugative elements: unraveling the SetCD regulon

Nucleic Acids Res. 2015 Feb 27;43(4):2045-56. doi: 10.1093/nar/gkv071. Epub 2015 Feb 6.

Abstract

Integrative and conjugative elements (ICEs) of the SXT/R391 family have been recognized as key drivers of antibiotic resistance dissemination in the seventh-pandemic lineage of Vibrio cholerae. SXT/R391 ICEs propagate by conjugation and integrate site-specifically into the chromosome of a wide range of environmental and clinical Gammaproteobacteria. SXT/R391 ICEs bear setC and setD, two conserved genes coding for a transcriptional activator complex that is essential for activation of conjugative transfer. We used chromatin immunoprecipitation coupled with exonuclease digestion (ChIP-exo) and RNA sequencing (RNA-seq) to characterize the SetCD regulon of three representative members of the SXT/R391 family. We also identified the DNA sequences bound by SetCD in MGIVflInd1, a mobilizable genomic island phylogenetically unrelated to SXT/R391 ICEs that hijacks the conjugative machinery of these ICEs to drive its own transfer. SetCD was found to bind a 19-bp sequence that is consistently located near the promoter -35 element of SetCD-activated genes, a position typical of class II transcriptional activators. Furthermore, we refined our understanding of the regulation of excision from and integration into the chromosome for SXT/R391 ICEs and demonstrated that de novo expression of SetCD is crucial to allow integration of the incoming ICE DNA into a naive host following conjugative transfer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Conjugation, Genetic*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / metabolism*
  • Gene Expression Regulation, Bacterial*
  • Genomic Islands
  • Mutation
  • Nucleotide Motifs
  • Operator Regions, Genetic
  • Promoter Regions, Genetic
  • Regulon*
  • Trans-Activators / genetics
  • Trans-Activators / metabolism*
  • Transcriptional Activation

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • Trans-Activators