title: IcgA is a virulence factor of Rhodococcus equi that modulates intracellular growth.
abstract: Virulence of the intracellular pathogen Rhodococcus equi depends on a 21.3-kb pathogenicity island located on a conjugative plasmid. To date, the only nonregulatory pathogenicity island-encoded virulence factor identified is the cell envelope-associated VapA protein. Although the pathogenicity islands from porcine and equine R. equi isolates have undergone major rearrangements, the virR operon (virR-icgA-vapH-orf7-virS) is highly conserved in both, suggesting these genes play an important role in pathogenicity. VirR and VirS are transcriptional regulators controlling expression of pathogenicity island genes, including vapA. Here, we show that while vapH and orf7 are dispensable for intracellular growth of R. equi, deletion of icgA, formerly known as orf5, encoding a major facilitator superfamily transport protein, elicited an enhanced growth phenotype in macrophages and a significant reduction in macrophage viability, while extracellular growth in broth remained unaffected. Transcription of virS, located downstream of icgA, and vapA was not affected by the icgA deletion during growth in broth or in macrophages, showing that the enhanced growth phenotype caused by deletion of icgA was not mediated through abnormal transcription of these genes. Transcription of icgA increased 6-fold within 2 h following infection of macrophages and remained significantly higher 48 h postinfection compared to levels at the start of the infection. The major facilitator superfamily transport protein IcgA is the first factor identified in R. equi that negatively affects intracellular replication. Aside from VapA, it is only the second pathogenicity island-encoded structural protein shown to play a direct role in intracellular growth of this pathogenic actinomycete.
authors.affiliation: UCD School of Biomolecular and Biomedical Science and UCD Conway Institute, University College Dublin, Dublin, Ireland.
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Wang
authors.forename: Garry B
authors.initials: GB
authors.lastname: Coulson
authors.forename: Aleksandra A
authors.initials: AA
authors.lastname: Miranda-Casoluengo
authors.forename: Raúl
authors.initials: R
authors.lastname: Miranda-Casoluengo
authors.forename: Mary K
authors.initials: MK
authors.lastname: Hondalus
authors.forename: Wim G
authors.initials: WG
authors.lastname: Meijer
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D016929:Rhodococcus equi / Q000378:metabolism* / Q000502:physiology*; D059467:Transcriptome; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D037521:Virulence Factors