Clade-1 Vap virulence proteins of Rhodococcus equi are associated with the cell surface and support intracellular growth in macrophages

PLoS One. 2025 Jan 6;20(1):e0316541. doi: 10.1371/journal.pone.0316541. eCollection 2025.

Abstract

The multi-host pathogen Rhodococcus equi is a parasite of macrophages preventing maturation of the phagolysosome, thus creating a hospitable environment supporting intracellular growth. Virulent R. equi isolated from foals, pigs and cattle harbor a host-specific virulence plasmid, pVAPA, pVAPB and pVAPN respectively, which encode a family of 17 Vap proteins belonging to seven monophyletic clades. We examined all 17 Vap proteins for their ability to complement intracellular growth of a R. equi ΔvapA strain, and show that only vapK1, vapK2 and vapN support growth in murine macrophages of this strain. We show that only the clade-1 proteins VapA, VapK1, VapK2 and VapN are located on the R. equi cell surface. The pVAPB plasmid encodes three clade-1 proteins: VapK1, VapK2 and VapB. The latter was not able to support intracellular growth and was not located on the cell surface. We previously showed that the unordered N-terminal VapA sequence is involved in cell surface localisation of VapA. We here show that although the unordered N-terminus of the 17 Vap proteins is highly variable in length and sequence, it is conserved within clades, which is consistent with our observation that the N-terminus of clade-1 Vap proteins plays a role in cell surface localisation.

MeSH terms

  • Actinomycetales Infections / microbiology
  • Animals
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / metabolism
  • Cell Membrane / metabolism
  • Macrophages* / microbiology
  • Mice
  • Rhodococcus equi* / genetics
  • Rhodococcus equi* / growth & development
  • Rhodococcus equi* / metabolism
  • Rhodococcus equi* / pathogenicity
  • Virulence
  • Virulence Factors / genetics
  • Virulence Factors / metabolism

Substances

  • Bacterial Proteins
  • Virulence Factors
  • VapA protein, Rhodococcus equi

Grants and funding

This research was funded by grants from University College Dublin (WGM) and the China Scholarship Council (YT&CC), and by a grant from the Tübitak-2219 programme, the Scientific and Technological Research Council of Türkiye (ZY). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript”.