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Nucleic Acids Research, 2000, Vol. 28, No. 18 3666-3673
© 2000 Oxford University Press

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

Jong-Wook Kim, Hyung-Cheol Kim1, Gyoung-Mi Kim, Jun-Mo Yang1,2, Jef D. Boeke3 and Kiebang Nam1,*

Department of Biochemistry, Inha University College of Medicine, Inchon, Republic of Korea, 1Clinical Research Center, Samsung Biomedical Research Institute, 50 Ilwon Dong, Kangnam Ku, Seoul 135-230, Republic of Korea, 2Department of Dermatology, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea, 3Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD, USA

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.

* To whom correspondence should be addressed at present address: Laboratory of Eukaryotic Gene Regulation, National Institute of Child Health and Human Development, NIH, Bethesda, MD 20892, USA. Tel: +1 301 435 5762; Fax: +1 301 496 4491; Email: knam@box-k.nih.gov


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