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

Biochem Biophys Res Commun. 2004 Nov 12;324(2):719-28. doi: 10.1016/j.bbrc.2004.09.112.


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 14alpha-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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Algorithms
  • Amino Acid Sequence
  • Antifungal Agents / pharmacology
  • Cloning, Molecular
  • Cryptococcus neoformans / enzymology*
  • Cryptococcus neoformans / genetics*
  • Cytochrome P-450 Enzyme System / chemistry*
  • Cytochrome P-450 Enzyme System / genetics*
  • DNA Primers / chemistry
  • DNA, Complementary / metabolism
  • Genes, Fungal
  • Genetic Complementation Test
  • Introns
  • Molecular Sequence Data
  • Mutation
  • Oxidoreductases / chemistry*
  • Oxidoreductases / genetics*
  • Phylogeny
  • Plasmids / metabolism
  • Polymerase Chain Reaction
  • RNA / metabolism
  • Saccharomyces cerevisiae / metabolism
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Sterol 14-Demethylase


  • Antifungal Agents
  • CYP51A1 protein, human
  • DNA Primers
  • DNA, Complementary
  • RNA
  • Cytochrome P-450 Enzyme System
  • Oxidoreductases
  • Sterol 14-Demethylase