Ehrlichia chaffeensis TRP47 enters the nucleus via a MYND-binding domain-dependent mechanism and predominantly binds enhancers of host genes associated with signal transduction, cytoskeletal organization, and immune response

PLoS One. 2018 Nov 8;13(11):e0205983. doi: 10.1371/journal.pone.0205983. eCollection 2018.

Abstract

Ehrlichia chaffeensis is an obligately intracellular bacterium that establishes infection in mononuclear phagocytes through largely undefined reprogramming strategies including modulation of host gene transcription. In this study, we demonstrate that the E. chaffeensis effector TRP47 enters the host cell nucleus and binds regulatory regions of host genes relevant to infection. TRP47 was observed in the nucleus of E. chaffeensis-infected host cells, and nuclear localization was dependent on a variant MYND-binding domain. An electrophoretic mobility shift assay (EMSA) demonstrated that TRP47 directly binds host DNA via its tandem repeat domain. Utilizing chromatin immunoprecipitation followed by high-throughput DNA sequencing (ChIP-seq) with E. chaffeensis-infected cells, TRP47 was found to bind at multiple sites in the human genome (n = 2,051 at p < 10-30). Ontology analysis identified genes involved in functions such as immune response, cytoskeletal organization, and signal transduction. TRP47-bound genes included RNA-coding genes, many of these linked to cell proliferation or apoptosis. Comparison of TRP47 binding sites with those of previously-identified E. chaffeensis nucleomodulins identified multiple genes and gene functional categories in common including intracellular transport, cell signaling, and transcriptional regulation. Further, motif analysis followed by EMSA with synthetic oligonucleotides containing discovered motifs revealed a conserved TRP47 DNA-binding motif. This study reveals that TRP47 is a nucleomodulin that enters the nucleus via a MYND-binding domain and appears to play a role in host cell reprogramming by regulation of transcription.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Apoptosis / genetics
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics*
  • Binding Sites / genetics
  • Cell Nucleus / genetics
  • Cell Proliferation / genetics
  • Cytoskeleton / genetics
  • DNA-Binding Proteins / genetics*
  • Ehrlichia chaffeensis / genetics*
  • Ehrlichia chaffeensis / pathogenicity
  • Gene Expression Regulation
  • Genome, Human / genetics*
  • High-Throughput Nucleotide Sequencing
  • Host-Pathogen Interactions / genetics*
  • Humans
  • Immunity, Cellular / genetics
  • MYND Domains / genetics
  • Nucleotide Motifs / genetics
  • Protein Binding
  • Signal Transduction / genetics

Substances

  • Bacterial Proteins
  • DNA-Binding Proteins