Cloning and characterization of the lanosterol 14α-demethylase (ERG11) gene in Cryptococcus neoformans

S. G. Revankar, J. Fu, M. G. Rinaldi, S. L. Kelly, D. E. Kelly, D. C. Lamb, S. M. Keller, B. L. Wickes

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

The ergosterol pathway in fungal pathogens is an attractive antimicrobial target because it is unique from the major sterol (cholesterol) producing pathway in humans. Lanosterol 14α-demethylase is the target for a major class of antifungals, the azoles. In this study we have isolated the gene for this enzyme from Cryptococcus neoformans. The gene, ERG11, was recovered using degenerate PCR with primers designed with a novel algorithm called CODEHOP. Sequence analysis of Erg11p identified a highly conserved region typical of the cytochrome P450 class of mono-oxygenases. The gene was present in single copy in the genome and mapped to one end of the largest chromosome. Comparison of the protein sequence to a number of major human fungal pathogen Erg11p homologs revealed that the C. neoformans protein was highly conserved, and most closely related to the Erg11p homologs from other basidiomycetes. Functional studies demonstrated that the gene could complement a Saccharomyces cerevisiae erg11 mutant, which confirmed the identity of the C. neoformans gene.

Original languageEnglish (US)
Pages (from-to)719-728
Number of pages10
JournalBiochemical and Biophysical Research Communications
Volume324
Issue number2
DOIs
StatePublished - Nov 12 2004

Keywords

  • Candida
  • Coccidioides
  • Codehop
  • Histoplasma
  • Lanosterol demethylase

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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