The Yarrowia lipolytica LEU2 gene

Abstract

A 2810 bp DNA fragment containing the beta-isopropylmalate dehydrogenase gene of the dimorphic yeast Yarrowia lipolytica has been sequenced. The sequence contains an open reading frame of 405 codons, predicting a protein of 43,366 molecular weight. Protein sequence homology with the polypeptide encoded by the LEU2 gene of Saccharomyces cerevisiae is 64%, whereas DNA sequence homology is 61%. The 5'- and 3'-flanking regions of the Y. lipolytica LEU2 gene share only some general structural features common to genes of S. cerevisiae such as the presence and location of TATA boxes, CAAT boxes, CACACA repeats, the lack of G residues in the 5'-untranslated region and 3'-transcription terminators. Transcription of a 1.4 kb mRNA begins at a small cluster of sites approximately 40 base pairs before the initial ATG.

Leucine Biosynthesis in Fungi: Entering Metabolism through the Back Door

“…This gene is therefore one of the best-studied genes of the branched-chain amino acid pathway in fungi. A partial list of organisms from which a LEU2 homologue has been cloned, sequenced, or otherwise characterized includes (in addition to the C. maltosa example mentioned above) the yeast Yarrowia lipolytica (29), the methylotrophic yeasts Hansenula polymorpha (2) and Pichia anomala (30), and the industrially used Kluyveromyces marxianus (12) and Aspergillus niger (111). A. niger elaborates two isoenzymes of ␤-isopropylmalate dehydrogenase (Leu2A and Leu2B) that are only 52% identical at the amino acid level.…”

The extracellular acid protease gene of Yarrowia lipolytica: sequence and pH-regulated transcription

“…lipobtica genes, XPR2 (Davidow et al, 1987a), LEU2 (Davidow et al, 1987b) and PYK7 (Strick e t al., 1992). There is very good agreement in terms of preferred codon usage throughout.…”

Further stabilization of 3-isopropylmalate dehydrogenase of an extreme thermophile, Thermus thermophilus, by a suppressor mutation method

“…This enzyme is on the leucine biosynthetic pathway. The leuB gene encoding IPMDH has been cloned and sequenced from a variety of microorganisms such as yeasts (2,6,7,26,34), bacilli (11,29,30). Salmonella typhimurium (3), a cyanobacterium, Spirulina platensis (4), and two extreme thermophiles, Thermus thermophilus (12,14) and Thermus aquaticus (15).…”