Molecular and functional analysis of the LYS1 gene of Candida albicans

Abstract

The LYS1 gene of Candida albicans has been localized to a 1.8-kb DNA fragment present on the plasmid YpBRG2. YpBRG2 has been shown to complement the saccharopine dehydrogenase mutant Stx4-4A of Saccharomyces cerevisiae. Transformants of S. cerevisiae Stx4-4A exhibited significant saccharopine dehydrogenase activity, and cells that had lost YpBRG2 after nonselective growth had no enzyme activity. The DNA sequence of the LYS1 gene has been determined. The LYS1 DNA contains typical yeast upstream regulatory sequences, including the GCN4 motif and candidate sequences responsible for transcription termination within the 3' noncoding region. The fragment contained an open reading frame of 1,146 nucleotides coding for a putative protein of 382 amino acids. The open reading frame has 60% identity at the nucleotide level and 71% similarity at the amino acid level to the LYS5 gene of Yarrowia lipolytica, which is believed to code for saccharopine dehydrogenase. A peptide of 11 amino acids has been found, which is present in S. cerevisiae, Y. lipolytica, and C. albicans. This peptide can be expanded to 16 amino acids when the sequences from Y. lipolytica and C. albicans are compared. A motif responsible for the binding of the adenosine residue of NADH has been described previously and is very similar to this peptide, which may be the site of NADH binding in the saccharopine dehydrogenase of C. albicans.