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

Jong Wook Kim, Hyung Cheol Kim, Gyoung Mi Kim, Jun Mo Yang, Jef D. Boeke, Kiebang Nam

    Research output: Contribution to journalArticle

    Abstract

    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, Schizosacharomyces 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.

    Original languageEnglish (US)
    Pages (from-to)3666-3673
    Number of pages8
    JournalNucleic acids research
    Volume28
    Issue number18
    DOIs
    StatePublished - Sep 15 2000

    ASJC Scopus subject areas

    • Genetics

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