Yeast lariat debranching enzyme

Substrate and sequence specificity

Kiebang Nam, Robert H E Hudson, Karen B. Chapman, Kanjana Ganeshan, Masad J. Damha, Jef D. Boeke

Research output: Contribution to journalArticle

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 - Aug 12 1994

Fingerprint

Substrate Specificity
Yeast
Yeasts
RNA
Substrates
Enzymes
Purines
Introns
Nucleic Acids
Assays
lariat debranching enzyme
msDNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Nam, K., Hudson, R. H. E., Chapman, K. B., Ganeshan, K., Damha, M. J., & Boeke, J. D. (1994). Yeast lariat debranching enzyme: Substrate and sequence specificity. Journal of Biological Chemistry, 269(32), 20613-20621.

Yeast lariat debranching enzyme : Substrate and sequence specificity. / Nam, Kiebang; Hudson, Robert H E; Chapman, Karen B.; Ganeshan, Kanjana; Damha, Masad J.; Boeke, Jef D.

In: Journal of Biological Chemistry, Vol. 269, No. 32, 12.08.1994, p. 20613-20621.

Research output: Contribution to journalArticle

Nam, K, Hudson, RHE, Chapman, KB, Ganeshan, K, Damha, MJ & Boeke, JD 1994, 'Yeast lariat debranching enzyme: Substrate and sequence specificity', Journal of Biological Chemistry, vol. 269, no. 32, pp. 20613-20621.
Nam K, Hudson RHE, Chapman KB, Ganeshan K, Damha MJ, Boeke JD. Yeast lariat debranching enzyme: Substrate and sequence specificity. Journal of Biological Chemistry. 1994 Aug 12;269(32):20613-20621.
Nam, Kiebang ; Hudson, Robert H E ; Chapman, Karen B. ; Ganeshan, Kanjana ; Damha, Masad J. ; Boeke, Jef D. / Yeast lariat debranching enzyme : Substrate and sequence specificity. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 32. pp. 20613-20621.
@article{45d52a59627942b1b22e0cd11cb17953,
title = "Yeast lariat debranching enzyme: Substrate and sequence specificity",
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.",
author = "Kiebang Nam and Hudson, {Robert H E} and Chapman, {Karen B.} and Kanjana Ganeshan and Damha, {Masad J.} and Boeke, {Jef D.}",
year = "1994",
month = "8",
day = "12",
language = "English (US)",
volume = "269",
pages = "20613--20621",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "32",

}

TY - JOUR

T1 - Yeast lariat debranching enzyme

T2 - Substrate and sequence specificity

AU - Nam, Kiebang

AU - Hudson, Robert H E

AU - Chapman, Karen B.

AU - Ganeshan, Kanjana

AU - Damha, Masad J.

AU - Boeke, Jef D.

PY - 1994/8/12

Y1 - 1994/8/12

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0028148071&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028148071&partnerID=8YFLogxK

M3 - Article

VL - 269

SP - 20613

EP - 20621

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 32

ER -