Skip Navigation

This Article
Right arrow Print PDF (3500K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (33)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Zabel, P.
Right arrow Articles by Van Kammen, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zabel, P.
Right arrow Articles by Van Kammen, A.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1981, Vol. 9, No. 11 2433-2453
© 1981

ENZYMOLOGY

Terminal uridylyl transferase of Vigna unguiculata: purification and characterization of an enzyme catalyzing the addition of a single UMP residue to the 3'-end of an RNA primer

Pim Zabel, Lambert Dorssers, Karel Wernars and Albert Van Kammen

Department of Molecular Biology, Agricultural University De Dreijen 11, 6703 BC Wageningen, The Netherlands

Received April 16, 1981. An enzyme which catalyzes the addition of a single UMP residue from UTP to the 3'-end of an RNA primer and which is referred to as terminal uridylyl transferase (TUT) has been extensively purified from the membrane fraction of Vigna unguiculata leaves. The purification procedure involved (i) solubilization by cation depletion (ii) DEAE-Sepharose CL-6B column chromatography (iii) affinity chromatography on poly(U)-Sepharose 4B and (iv) glycerol gradient centrifugation. The molecular weight of the native enzyme was approximately 50,000 as determined by velocity sedimentation. Under conditions that were optimal for UMP-incorporation (5 mM Mg2+, low salt, 30°C) TUT displayed a marked specificity for UTP as substrate, was unable to incorporate deoxyribonucleoside triphosphates and required a single-stranded oligo- or polyribonucleotide as primer. When oligoA20. tRNAasP of E.coli or alfalfa mosaic virus RNA 4 were used as primers at various substrate to primer ratio's, the vast majority of the product appeared to consist of primer molecules elongated with a single UMP residue as shown by polyacrylamide gelelectrophoresis and nearest neighbour analysis. We believe TUT to be a novel enzyme which has not been reported before and which may be a feasible tool in RNA sequencing as it enables the specific 3'-terminal labeling of RNA molecules.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 RNAHome page
R. Trippe, E. Guschina, M. Hossbach, H. Urlaub, R. Luhrmann, and B.-J. Benecke
Identification, cloning, and functional analysis of the human U6 snRNA-specific terminal uridylyl transferase
RNA, August 1, 2006; 12(8): 1494 - 1504.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
I. Aphasizheva, R. Aphasizhev, and L. Simpson
RNA-editing Terminal Uridylyl Transferase 1: IDENTIFICATION OF FUNCTIONAL DOMAINS BY MUTATIONAL ANALYSIS
J. Biol. Chem., June 4, 2004; 279(23): 24123 - 24130.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
M. T. McManus, B. K. Adler, V. W. Pollard, and S. L. Hajduk
Trypanosoma brucei Guide RNA Poly(U) Tail Formation Is Stabilized by Cognate mRNA
Mol. Cell. Biol., February 1, 2000; 20(3): 883 - 891.
[Abstract] [Full Text]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.