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Nucleotide sequence and chromosomal localization of the gene encoding the old yellow enzyme from Kluyveromyces lactis
Author(s) -
Miranda Manuel,
Ramírez Jorge,
Guevara Soledad,
OngayLarios Laura,
Peña Antonio,
Coria Roberto
Publication year - 1995
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320110509
Subject(s) - biology , kluyveromyces lactis , microbiology and biotechnology , nucleic acid sequence , saccharomyces cerevisiae , biochemistry , peptide sequence , open reading frame , gene , kluyveromyces marxianus , kluyveromyces , genetics
A 6·6 kb genomic DNA fragment from the yeast Kluyveromyces lactis was isolated. Sequence analysis of this fragment revealed the presence of two incomplete open reading frames (ORFs) in one strand, one coding for the carboxyl terminus of the plasma membrane H + ‐ATPase and the other for the amino terminus of an unidentified product. In the complementary strand, a full‐length ORF which encodes for a protein homologous to the yeast NADPH‐dependent Old Yellow Enzyme was found. The deduced amino acid sequence of this ORF predicts a protein of 398 residues with 84% similarity in its full length to OYE1 from Saccharomyces carlsbergensis and OYE2 from Saccharomyces cerevisiae . In addition, an internal region showed considerable similarity to the bile acid‐inducible polypeptide from Eubacterium sp. , to the NADH oxidase from Thermoanaerobium brockii , to the trimethylamino dehydrogenase from bacterium W 3 A 1 and to the estrogen‐binding protein from Candida albicans , suggesting a functional or structural relationship between them. Inactivation of the KYE1 ( K luyveromyces Y ellow E nzyme) gene by deletion of 0·6 kb fragment between positions +358 and +936 produced viable cells with a slight increase in their generation time. Haploid cells carrying the disrupted allele showed one‐third of the NADPH oxidase activity, compared to wild‐type cells. Southern blotting analysis of digested DNA and chromosomes separated by contour‐clamped homogeneous electric field electrophoresis from K. lactis indicated that this is a single‐copy gene and it is localized on chromosome II, whose molecular size has been estimated to be approximately 1·3 Mb. The sequence reported in this paper has been deposited in the GenBank data base (Accession No. L37452).