Human RNA lariat debranching enzyme cDNA complements the phenotypes of Saccharomyces cerevisiae dbr1 and Schizosaccharomyces pombe dbr1 mutants

• Published in: Nucleic acids research Vol. 28; no. 18; pp. 3666 - 3673
• Main Authors: Kim, J W, Kim, H C, Kim, G M, Yang, J M, Boeke, J D, Nam, K
• Format: Journal Article
• Language: English
• Published: England Oxford Publishing Limited (England) 15.09.2000; Oxford University Press
• Subjects: Amino Acid Sequence; Cloning, Molecular; DBR1 gene; DNA, Complementary; DNA, Fungal; Escherichia coli; Expressed Sequence Tags; Genetic Complementation Test; HeLa Cells; Humans; Introns; Molecular Sequence Data; Mutation; Phenotype; RNA lariat debranching enzyme; RNA Nucleotidyltransferases - genetics; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Schizosaccharomyces - genetics; Schizosaccharomyces pombe; Sequence Homology, Amino Acid
• ISSN: 1362-4962; 0305-1048; 1362-4962
• DOI: 10.1093/nar/28.18.3666

The cDNA encoding the human RNA lariat debranching enzyme (hDBR1) was identified and cloned by searching the Expressed Sequence Tag (EST) database and screening a HeLa cDNA library, based on predicted amino acid sequence homologies with the Saccharomyces cerevisiae, Schizosaccharomyces pombe and Caenorhabditis elegans debranching enzymes. The hDBR1 cDNA expressed in Escherichia coli showed debranching activity in vitro and was also shown to be functional in an interspecies specific complementation experiment. hDBR1 cDNA in a S. cerevisiae expression vector complemented the intron accumulation phenotype of a S. cerevisiae dbr1 null mutant. Integration of the cDNA for hDBR1 into the ura4 locus of S. pombe also complemented both the intron accumulation and slow growth phenotypes of a S. pombe dbr1 null mutant strain. Comparison of the amino acid sequence of hDBR1 with the other DBR protein sequences showed several conserved regions, with 40, 44 and 43% identity to the S. cerevisiae, S. pombe and C. elegans debranching enzymes, respectively.