Expression of Paracoccidioides brasiliensis CHS3 in a Saccharomyces cerevisiae chs3 null mutant demonstrates its functionality as a chitin synthase gene

Yeast. 2010 May;27(5):293-300. doi: 10.1002/yea.1748.

Abstract

We report the isolation and sequencing of CHS3, a gene that encodes one of several chitin synthases in Paracoccidioides brasiliensis, a medically important fungus restricted geographically to Latin America. The gene contains a single open reading frame of 3817 bp with two introns (71 and 86 bp) and encodes a 1220 amino acid polypeptide with high similarity to other fungal chitin synthases. Northern analysis reveals a high expression of CHS3 in the pathogenic yeast-like phase of the fungus and at the end of the mycelium-yeast transition. Expression of P. brasiliensis CHS3 in a Saccharomyces cerevisiae chs3 null mutant enhanced calcofluor white staining in parallel to an increase in total chitin synthase activity and chitin content in its cell wall.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Wall / chemistry
  • Chitin / analysis
  • Chitin / biosynthesis
  • Chitin Synthase / chemistry
  • Chitin Synthase / genetics*
  • Chitin Synthase / metabolism
  • Cloning, Molecular
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics*
  • Fungal Proteins / metabolism
  • Genes, Fungal
  • Molecular Sequence Data
  • Mutation
  • Open Reading Frames
  • Paracoccidioides / enzymology*
  • Paracoccidioides / genetics*
  • Paracoccidioides / growth & development
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Transformation, Genetic

Substances

  • Fungal Proteins
  • Chitin
  • Chitin Synthase

Associated data

  • GENBANK/AF107623