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Reference: Nam K, et al. (1994) Yeast lariat debranching enzyme. Substrate and sequence specificity. J Biol Chem 269(32):20613-21

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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.

Reference Type
Journal Article
Authors
Nam K, Hudson RH, Chapman KB, Ganeshan K, Damha MJ, Boeke JD
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