Yeast lariat debranching enzyme. Substrate and sequence specificity

K. Nam, R. H.E. Hudson, K. B. Chapman, K. Ganeshan, M. J. Damha, J. D. Boeke

Research output: Contribution to journalArticlepeer-review

Abstract

Yeast RNA lariat debranching enzyme has been purified to near homogeneity using a bacterial overproducer of the enzyme. The enzyme is capable of digesting a variety of branched nucleic acid substrates, including group II intron lariats, multicopy single-stranded DNAs (msDNAs), and a variety of synthetic branched RNAs. A trinucleotide release assay using radiolabeled msDNA substrates was developed and used to determine the basic biochemical parameters for the enzyme. The debranching enzyme shows a strong preference for purines at the 2'-position in both msDNA and synthetic branched RNA substrates, in accord with the structure of its native substrate, which always has a 2'-G residue. The use of small synthetic branched RNA substrates will allow systematic mechanistic and structural studies of this unique enzyme.

Original languageEnglish (US)
Pages (from-to)20613-20621
Number of pages9
JournalJournal of Biological Chemistry
Volume269
Issue number32
StatePublished - 1994

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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