Evidence for a Pneumocystis carinii Flo8-like transcription factor

insights into organism adhesion

Theodore J. Kottom, Andrew Harold Limper

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Pneumocystis carinii (Pc) adhesion to alveolar epithelial cells is well established and is thought to be a prerequisite for the initiation of Pneumocystis pneumonia. Pc binding events occur in part through the major Pc surface glycoprotein Msg, as well as an integrin-like molecule termed PcInt1. Recent data from the Pc sequencing project also demonstrate DNA sequences homologous to other genes important in Candida spp. binding to mammalian host cells, as well as organism binding to polystyrene surfaces and in biofilm formation. One of these genes, flo8, a transcription factor needed for downstream cAMP/PKA-pathway-mediated activation of the major adhesion/flocculin Flo11 in yeast, was cloned from a Pc cDNA library utilizing a partial sequence available in the Pc genome database. A CHEF blot of Pc genomic DNA yielded a single band providing evidence this gene is present in the organism. BLASTP analysis of the predicted protein demonstrated 41 % homology to the Saccharomyces cerevisiae Flo8. Northern blotting demonstrated greatest expression at pH 6.0–8.0, pH comparable to reported fungal biofilm milieu. Western blot and immunoprecipitation assays of PcFlo8 protein in isolated cyst and tropic life forms confirmed the presence of the cognate protein in these Pc life forms. Heterologous expression of Pcflo8 cDNA in flo8Δ-deficient yeast strains demonstrated that the Pcflo8 was able to restore yeast binding to polystyrene and invasive growth of yeast flo8Δ cells. Furthermore, Pcflo8 promoted yeast binding to HEK293 human epithelial cells, strengthening its functional classification as a Flo8 transcription factor. Taken together, these data suggest that PcFlo8 is expressed by Pc and may exert activity in organism adhesion and biofilm formation.

Original languageEnglish (US)
Pages (from-to)73-84
Number of pages12
JournalMedical Microbiology and Immunology
Volume205
Issue number1
DOIs
StatePublished - Feb 1 2016

Fingerprint

Pneumocystis carinii
Transcription Factors
Yeasts
Biofilms
Polystyrenes
Genes
Alveolar Epithelial Cells
Proteins
Pneumocystis Pneumonia
Gene Library
Candida
Immunoprecipitation
Integrins
Northern Blotting
Saccharomyces cerevisiae
Cysts
Complementary DNA
Western Blotting
Epithelial Cells
Genome

Keywords

  • Cell adhesion
  • Flo8
  • Pathogenesis
  • Pneumocystis

ASJC Scopus subject areas

  • Immunology and Allergy
  • Microbiology (medical)
  • Immunology

Cite this

Evidence for a Pneumocystis carinii Flo8-like transcription factor : insights into organism adhesion. / Kottom, Theodore J.; Limper, Andrew Harold.

In: Medical Microbiology and Immunology, Vol. 205, No. 1, 01.02.2016, p. 73-84.

Research output: Contribution to journalArticle

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