1887

Abstract

The virulence plasmid of the pathogenic actinomycete is essential for proliferation of this pathogen in macrophages and the development of disease. The pathogenicity island of this plasmid encodes a family of virulence-associated proteins (Vap), one of which (VapA) is a virulence factor. This paper describes the operon ( o-expressed ene), located upstream of the operon. Transcription of the operon gave rise to transcripts with a half-life similar to those determined for other virulence plasmid genes (1.8 min). Transcription started at a promoter similar to the promoter, and proceeded through an inefficient terminator into the downstream gene. In addition, is also transcribed from a promoter downstream of The and operons were coordinately regulated by temperature and pH in a synergistic manner. The latter parameter only affected transcription at higher growth temperatures, indicating that temperature is the dominant regulatory signal. Transcription of the operon increased 10-fold during the late exponential and stationary growth phases. Transcription was also upregulated during the initial hours following phagocytosis by phagocytic cells. In contrast to and , the gene is conserved in the porcine VapB-encoding plasmid, as well as in pathogenic mycobacteria. The coordinated regulation of and , transcription of following phagocytosis and conservation of in pathogenic mycobacteria indicate a role for and the genes in the virulence of .

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2011-08-01
2019-11-18
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References

  1. Butcher P. D. , Mangan J. A. , Monahan I. M. . ( 1998; ). Intracellular gene expression. Analysis of RNA from mycobacteria in macrophages using RT-PCR. . Methods Mol Biol 101:, 285–306.[PubMed].
    [Google Scholar]
  2. Buttner M. J. , Chater K. F. , Bibb M. J. . ( 1990; ). Cloning, disruption, and transcriptional analysis of three RNA polymerase sigma factor genes of Streptomyces coelicolor A3(2). . J Bacteriol 172:, 3367–3378.[PubMed].
    [Google Scholar]
  3. Byrne B. A. , Prescott J. F. , Palmer G. H. , Takai S. , Nicholson V. M. , Alperin D. C. , Hines S. A. . ( 2001; ). Virulence plasmid of Rhodococcus equi contains inducible gene family encoding secreted proteins. . Infect Immun 69:, 650–656. [CrossRef].[PubMed].
    [Google Scholar]
  4. Byrne G. A. , Russell D. A. , Chen X. , Meijer W. G. . ( 2007; ). Transcriptional regulation of the virR operon of the intracellular pathogen Rhodococcus equi . . J Bacteriol 189:, 5082–5089. [CrossRef].[PubMed].
    [Google Scholar]
  5. Byrne G. A. , Boland C. A. , O’Connell E. P. , Meijer W. G. . ( 2008; ). Differential mRNA stability of the vapAICD operon of the facultative intracellular pathogen Rhodococcus equi . . FEMS Microbiol Lett 280:, 89–94. [CrossRef].[PubMed].
    [Google Scholar]
  6. Cole S. T. , Brosch R. , Parkhill J. , Garnier T. , Churcher C. , Harris D. , Gordon S. V. , Eiglmeier K. , Gas S. et al. ( 1998; ). Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. . Nature 393:, 537–544. [CrossRef].[PubMed].
    [Google Scholar]
  7. De La Peña-Moctezuma A. , Prescott J. F. , Goodfellow M. . ( 1996; ). Attempts to find phenotypic markers of the virulence plasmid of Rhodococcus equi . . Can J Vet Res 60:, 29–33.[PubMed].
    [Google Scholar]
  8. Demangel C. , Stinear T. P. , Cole S. T. . ( 2009; ). Buruli ulcer: reductive evolution enhances pathogenicity of Mycobacterium ulcerans . . Nat Rev Microbiol 7:, 50–60. [CrossRef].[PubMed].
    [Google Scholar]
  9. Devulder G. , Pérouse de Montclos M. , Flandrois J. P. . ( 2005; ). A multigene approach to phylogenetic analysis using the genus Mycobacterium as a model. . Int J Syst Evol Microbiol 55:, 293–302. [CrossRef].[PubMed].
    [Google Scholar]
  10. Felsenstein J. . ( 1989; ). phylip – phylogeny inference package (version 3.2). . Cladistics 5:, 164–166.
    [Google Scholar]
  11. Fernandez-Mora E. , Polidori M. , Lührmann A. , Schaible U. E. , Haas A. . ( 2005; ). Maturation of Rhodococcus equi-containing vacuoles is arrested after completion of the early endosome stage. . Traffic 6:, 635–653. [CrossRef].[PubMed].
    [Google Scholar]
  12. Fleischmann R. D. , Alland D. , Eisen J. A. , Carpenter L. , White O. , Peterson J. , DeBoy R. , Dodson R. , Gwinn M. et al. ( 2002; ). Whole-genome comparison of Mycobacterium tuberculosis clinical and laboratory strains. . J Bacteriol 184:, 5479–5490. [CrossRef].[PubMed].
    [Google Scholar]
  13. Giguère S. , Hondalus M. K. , Yager J. A. , Darrah P. , Mosser D. M. , Prescott J. F. . ( 1999; ). Role of the 85-kilobase plasmid and plasmid-encoded virulence-associated protein A in intracellular survival and virulence of Rhodococcus equi . . Infect Immun 67:, 3548–3557.[PubMed].
    [Google Scholar]
  14. Hondalus M. K. , Mosser D. M. . ( 1994; ). Survival and replication of Rhodococcus equi in macrophages. . Infect Immun 62:, 4167–4175.[PubMed].
    [Google Scholar]
  15. Hondalus M. K. , Diamond M. S. , Rosenthal L. A. , Springer T. A. , Mosser D. M. . ( 1993; ). The intracellular bacterium Rhodococcus equi requires Mac-1 to bind to mammalian cells. . Infect Immun 61:, 2919–2929.[PubMed].
    [Google Scholar]
  16. Hovel-Miner G. , Pampou S. , Faucher S. P. , Clarke M. , Morozova I. , Morozov P. , Russo J. J. , Shuman H. A. , Kalachikov S. . ( 2009; ). σS controls multiple pathways associated with intracellular multiplication of Legionella pneumophila . . J Bacteriol 191:, 2461–2473. [CrossRef].[PubMed].
    [Google Scholar]
  17. Hughes K. L. , Sulaiman I. . ( 1987; ). The ecology of Rhodococcus equi and physicochemical influences on growth. . Vet Microbiol 14:, 241–250. [CrossRef].[PubMed].
    [Google Scholar]
  18. Iriarte M. , Stainier I. , Cornelis G. R. . ( 1995; ). The rpoS gene from Yersinia enterocolitica and its influence on expression of virulence factors. . Infect Immun 63:, 1840–1847.[PubMed].
    [Google Scholar]
  19. Jain S. , Bloom B. R. , Hondalus M. K. . ( 2003; ). Deletion of vapA encoding virulence associated protein A attenuates the intracellular actinomycete Rhodococcus equi . . Mol Microbiol 50:, 115–128. [CrossRef].[PubMed].
    [Google Scholar]
  20. Jones D. T. , Taylor W. R. , Thornton J. M. . ( 1992; ). The rapid generation of mutation data matrices from protein sequences. . Comput Appl Biosci 8:, 275–282.[PubMed].
    [Google Scholar]
  21. Kang J. G. , Hahn M. Y. , Ishihama A. , Roe J. H. . ( 1997; ). Identification of sigma factors for growth phase-related promoter selectivity of RNA polymerases from Streptomyces coelicolor A3(2). . Nucleic Acids Res 25:, 2566–2573. [CrossRef].[PubMed].
    [Google Scholar]
  22. Kelly B. G. , Wall D. M. , Boland C. A. , Meijer W. G. . ( 2002; ). Isocitrate lyase of the facultative intracellular pathogen Rhodococcus equi . . Microbiology 148:, 793–798.[PubMed].
    [Google Scholar]
  23. Klinkert B. , Narberhaus F. . ( 2009; ). Microbial thermosensors. . Cell Mol Life Sci 66:, 2661–2676. [CrossRef].[PubMed].
    [Google Scholar]
  24. Larkin M. A. , Blackshields G. , Brown N. P. , Chenna R. , McGettigan P. A. , McWilliam H. , Valentin F. , Wallace I. M. , Wilm A. et al. ( 2007; ). clustal_w and clustal_x version 2.0. . Bioinformatics 23:, 2947–2948. [CrossRef].[PubMed].
    [Google Scholar]
  25. Letek M. , Ocampo-Sosa A. A. , Sanders M. , Fogarty U. , Buckley T. , Leadon D. P. , González P. , Scortti M. , Meijer W. G. et al. ( 2008; ). Evolution of the Rhodococcus equi vap pathogenicity island seen through comparison of host-associated vapA and vapB virulence plasmids. . J Bacteriol 190:, 5797–5805. [CrossRef].[PubMed].
    [Google Scholar]
  26. Letek M. , González P. , Macarthur I. , Rodríguez H. , Freeman T. C. , Valero-Rello A. , Blanco M. , Buckley T. , Cherevach I. et al. ( 2010; ). The genome of a pathogenic Rhodococcus: cooptive virulence underpinned by key gene acquisitions. . PLoS Genet 6:, e1001145. [CrossRef].[PubMed].
    [Google Scholar]
  27. Li L. , Bannantine J. P. , Zhang Q. , Amonsin A. , May B. J. , Alt D. , Banerji N. , Kanjilal S. , Kapur V. . ( 2005; ). The complete genome sequence of Mycobacterium avium subspecies paratuberculosis . . Proc Natl Acad Sci U S A 102:, 12344–12349. [CrossRef].[PubMed].
    [Google Scholar]
  28. Lührmann A. , Mauder N. , Sydor T. , Fernandez-Mora E. , Schulze-Luehrmann J. , Takai S. , Haas A. . ( 2004; ). Necrotic death of Rhodococcus equi-infected macrophages is regulated by virulence-associated plasmids. . Infect Immun 72:, 853–862. [CrossRef].[PubMed].
    [Google Scholar]
  29. Meijer W. G. , Prescott J. F. . ( 2004; ). Rhodococcus equi . . Vet Res 35:, 383–396. [CrossRef].[PubMed].
    [Google Scholar]
  30. Miranda-CasoLuengo R. , Duffy P. S. , O’Connell E. P. , Graham B. J. , Mangan M. W. , Prescott J. F. , Meijer W. G. . ( 2005; ). The iron-regulated iupABC operon is required for saprophytic growth of the intracellular pathogen Rhodococcus equi at low iron concentrations. . J Bacteriol 187:, 3438–3444. [CrossRef].[PubMed].
    [Google Scholar]
  31. Muscatello G. , Leadon D. P. , Klayt M. , Ocampo-Sosa A. , Lewis D. A. , Fogarty U. , Buckley T. , Gilkerson J. R. , Meijer W. G. , Vazquez-Boland J. A. . ( 2007; ). Rhodococcus equi infection in foals: the science of ‘rattles’. . Equine Vet J 39:, 470–478. [CrossRef].[PubMed].
    [Google Scholar]
  32. Nielsen A. T. , Dolganov N. A. , Otto G. , Miller M. C. , Wu C. Y. , Schoolnik G. K. . ( 2006; ). RpoS controls the Vibrio cholerae mucosal escape response. . PLoS Pathog 2:, e109. [CrossRef].[PubMed].
    [Google Scholar]
  33. Ocampo-Sosa A. A. , Lewis D. A. , Navas J. , Quigley F. , Callejo R. , Scortti M. , Leadon D. P. , Fogarty U. , Vázquez-Boland J. A. . ( 2007; ). Molecular epidemiology of Rhodococcus equi based on traA, vapA, and vapB virulence plasmid markers. . J Infect Dis 196:, 763–769. [CrossRef].[PubMed].
    [Google Scholar]
  34. Pfaffl M. W. . ( 2004; ). Quantification strategies in real-time PCR. . In A–Z of Quantitative PCR, pp. 87–112. Edited by Bustin S. A. . . La Jolla, CA:: International University Line;.
    [Google Scholar]
  35. Platt T. . ( 1986; ). Transcription termination and the regulation of gene expression. . Annu Rev Biochem 55:, 339–372. [CrossRef].[PubMed].
    [Google Scholar]
  36. Polidori M. , Haas A. . ( 2006; ). VapI, a new member of the Rhodococcus equi Vap family. . Antonie van Leeuwenhoek 90:, 299–304. [CrossRef].[PubMed].
    [Google Scholar]
  37. Ren J. , Prescott J. F. . ( 2003; ). Analysis of virulence plasmid gene expression of intra-macrophage and in vitro grown Rhodococcus equi ATCC 33701. . Vet Microbiol 94:, 167–182. [CrossRef].[PubMed].
    [Google Scholar]
  38. Ren J. , Prescott J. F. . ( 2004; ). The effect of mutation on Rhodococcus equi virulence plasmid gene expression and mouse virulence. . Vet Microbiol 103:, 219–230. [CrossRef].[PubMed].
    [Google Scholar]
  39. Ripoll F. , Pasek S. , Schenowitz C. , Dossat C. , Barbe V. , Rottman M. , Macheras E. , Heym B. , Herrmann J. L. et al. ( 2009; ). Non mycobacterial virulence genes in the genome of the emerging pathogen Mycobacterium abscessus . . PLoS ONE 4:, e5660. [CrossRef].[PubMed].
    [Google Scholar]
  40. Rohde K. H. , Abramovitch R. B. , Russell D. G. . ( 2007; ). Mycobacterium tuberculosis invasion of macrophages: linking bacterial gene expression to environmental cues. . Cell Host Microbe 2:, 352–364. [CrossRef].[PubMed].
    [Google Scholar]
  41. Russell D. A. , Byrne G. A. , O’Connell E. P. , Boland C. A. , Meijer W. G. . ( 2004; ). The LysR-type transcriptional regulator VirR is required for expression of the virulence gene vapA of Rhodococcus equi ATCC 33701. . J Bacteriol 186:, 5576–5584. [CrossRef].[PubMed].
    [Google Scholar]
  42. Sambrook J. , Russell D. W. . ( 2001; ). Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;.
    [Google Scholar]
  43. Sekine M. , Tanikawa S. , Omata S. , Saito M. , Fujisawa T. , Tsukatani N. , Tajima T. , Sekigawa T. , Kosugi H. et al. ( 2006; ). Sequence analysis of three plasmids harboured in Rhodococcus erythropolis strain PR4. . Environ Microbiol 8:, 334–346. [CrossRef].[PubMed].
    [Google Scholar]
  44. Stinear T. P. , Seemann T. , Pidot S. , Frigui W. , Reysset G. , Garnier T. , Meurice G. , Simon D. , Bouchier C. et al. ( 2007; ). Reductive evolution and niche adaptation inferred from the genome of Mycobacterium ulcerans, the causative agent of Buruli ulcer. . Genome Res 17:, 192–200. [CrossRef].[PubMed].
    [Google Scholar]
  45. Stinear T. P. , Seemann T. , Harrison P. F. , Jenkin G. A. , Davies J. K. , Johnson P. D. , Abdellah Z. , Arrowsmith C. , Chillingworth T. et al. ( 2008; ). Insights from the complete genome sequence of Mycobacterium marinum on the evolution of Mycobacterium tuberculosis . . Genome Res 18:, 729–741. [CrossRef].[PubMed].
    [Google Scholar]
  46. Takai S. , Fukunaga N. , Kamisawa K. , Imai Y. , Sasaki Y. , Tsubaki S. . ( 1996; ). Expression of virulence-associated antigens of Rhodococcus equi is regulated by temperature and pH. . Microbiol Immunol 40:, 591–594.[PubMed].[CrossRef]
    [Google Scholar]
  47. Takai S. , Hines S. A. , Sekizaki T. , Nicholson V. M. , Alperin D. A. , Osaki M. , Takamatsu D. , Nakamura M. , Suzuki K. et al. ( 2000; ). DNA sequence and comparison of virulence plasmids from Rhodococcus equi ATCC 33701 and 103. . Infect Immun 68:, 6840–6847. [CrossRef].[PubMed].
    [Google Scholar]
  48. Toyooka K. , Takai S. , Kirikae T. . ( 2005; ). Rhodococcus equi can survive a phagolysosomal environment in macrophages by suppressing acidification of the phagolysosome. . J Med Microbiol 54:, 1007–1015. [CrossRef].[PubMed].
    [Google Scholar]
  49. von Bargen K. , Wohlmann J. , Taylor G. A. , Utermöhlen O. , Haas A. . ( 2011; ). Nitric oxide-mediated intracellular growth restriction of pathogenic Rhodococcus equi can be prevented by iron. . Infect Immun 79:, 2098–2111. [CrossRef].[PubMed].
    [Google Scholar]
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