RNA interference-mediated silencing of the YPS3 gene of Histoplasma capsulatum reveals virulence defects

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Megan L Bohse1, Jon P Woods

  • 1Department of Medical Microbiology and Immunology, 420 SMI, University of Wisconsin Medical School, 1300 University Avenue, Madison, WI 53706, USA.

Infection and Immunity
|April 4, 2007
PubMed

Insights

Silencing the YPS3 gene in Histoplasma capsulatum reduced fungal burdens in mouse organs, indicating YPS3

Area of Science:

  • Medical Mycology
  • Molecular Biology
  • Infectious Diseases

Background:

  • Histoplasma capsulatum (H. capsulatum) is a fungal pathogen causing histoplasmosis.
  • The YPS3 gene product is surface-localized and secreted during the yeast phase.
  • YPS3 expression varies with virulence levels.

Purpose of the Study:

  • To investigate the role of YPS3 in H. capsulatum pathogenesis.
  • To assess the in vitro and in vivo effects of YPS3 silencing.

Main Methods:

  • RNA interference (RNAi) was used to silence the YPS3 transcript.
  • Phenotypic analysis was performed in vitro and in a murine infection model.
  • Fungal burdens were quantified in organs of infected C57BL/6 mice.

Main Results:

  • YPS3-silenced mutants showed no in vitro growth defects at 37°C.
  • Mutants exhibited normal virulence in RAW 264.7 macrophage cell line.
  • Significantly decreased fungal burdens were observed in livers and spleens of infected mice.

Conclusions:

  • YPS3 is crucial for H. capsulatum colonization of host tissues, particularly in spleen and liver.
  • YPS3-mediated virulence appears to be dose-dependent and time-exacerbated.
  • Targeting YPS3 may offer a strategy to combat histoplasmosis.

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