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A DUF4148 family protein produced inside RAW264.7 cells is a critical Burkholderia pseudomallei virulence factor

Susan Welkos1, Irma Blanco1, Udoka Okaro1

  • 1Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases , Frederick, Maryland, USA.

Virulence
|August 25, 2020
PubMed

Insights

Burkholderia pseudomallei causes melioidosis. This study identified bacterial protein BPSS1996 essential for virulence and 274 host proteins involved in the immune response to infection.

Area of Science:

  • Microbiology
  • Proteomics
  • Immunology

Background:

  • * Burkholderia pseudomallei* is the causative agent of melioidosis, a serious infectious disease.
  • * B. pseudomallei* employs sophisticated mechanisms to survive within host phagocytic cells, including vacuole escape, cytosolic replication, and cell-to-cell spread.
  • * Understanding the molecular interactions between *B. pseudomallei* and host cells is crucial for developing effective treatments.

Purpose of the Study:

  • To identify bacterial proteins produced by *B. pseudomallei* within macrophages.
  • To identify host proteins modulated during *B. pseudomallei* infection.
  • To investigate the role of novel bacterial proteins in virulence.

Main Methods:

  • Proteomics analysis using liquid chromatography and tandem mass spectrometry on infected RAW264.7 murine macrophages.
  • Construction and virulence assessment of mutants in novel *B. pseudomallei* genes in BALB/c mice.
  • Indirect immunofluorescence to determine protein localization.

Main Results:

  • Approximately 160 bacterial proteins were identified, including known virulence factors and previously uncharacterized proteins like BPSS1996.
  • A mutant in *BPSS1996* exhibited significantly reduced virulence (55-fold higher LD50), indicating its essential role.
  • 274 host proteins, including chemokines and cytokines, were identified as differentially expressed in response to infection.

Conclusions:

  • *B. pseudomallei* protein BPSS1996 is critical for full virulence and is surface-localized.
  • The study provides insights into the host-pathogen interactions during melioidosis.
  • Identification of host response proteins may offer targets for therapeutic intervention.
Keywords:
Domain of unknown functionfacultative intracellular pathogenselect agent

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