Two Origins for the Gene Encoding α-Isopropylmalate Synthase in Fungi

BACKGROUND: The biosynthesis of leucine is a biochemical pathway common to prokaryotes, plants and fungi, but absent from humans and animals. The pathway is a proposed target for antimicrobial therapy. METHODOLOGY/PRINCIPAL FINDINGS: Here we identified the leuA gene encoding alpha-isopropylmalate synthase in the zygomycete fungus Phycomyces blakesleeanus using a genetic mapping approach with crosses between wild type and leucine auxotrophic strains. To confirm the function of the gene, Phycomyces leuA was used to complement the auxotrophic phenotype exhibited by mutation of the leu3+ gene of the ascomycete fungus Schizosaccharomyces pombe. Phylogenetic analysis revealed that the leuA gene in Phycomyces, other zygomycetes, and the chytrids is more closely related to homologs in plants and photosynthetic bacteria than ascomycetes or basidiomycetes, and suggests that the Dikarya have acquired the gene more recently. CONCLUSIONS/SIGNIFICANCE: The identification of leuA in Phycomyces adds to the growing body of evidence that some primary metabolic pathways or parts of them have arisen multiple times during the evolution of fungi, probably through horizontal gene transfer events

Cloning and expression analysis of beta-isopropylmalate dehydrogenase from potato

A full-length cDNA clone for β-isopropylmalate dehydrogenase from potato has been isolated and sequenced. The open reading frame is 1071 by in length encoding a protein of 357 amino acids which includes a 29 amino acid, putative chloroplastic transit peptide. The amino acid sequence shows 33.3% and 28.6% identity to β-isopropylmalate dehydrogenases from rape and Bacillus subtilis, respectively. Southern analysis shows that the gene is present in low copy number in potato, and in single copy in tomato and Arabidopsis. The gene is expressed in all tissues of the potato plant and its expression is increased by leucine, and leucine plus threonine, in contrast to the situation in yeast and prokaryotes. The gene is also induced by sucrose in a manner similar to that seen with genes involved in carbohydrate metabolism, which indicates that there may be some interaction at the transcriptional level between genes involved in carbon and nitrogen metabolism