Skip Navigation

This Article
Right arrow Print PDF (1380K)
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 (76)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Griffiths, A.D.
Right arrow Articles by Eperon, I.C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Griffiths, A.D.
Right arrow Articles by Eperon, I.C.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1987, Vol. 15, No. 10 4145-4162
© 1987

Articles

Stereospecificity of nucleases towards phosphorothioate-substituted RNA: stereochemistry of transcription by T7 RNA polymerase

A.D. Griffiths, B.V.L. Potter* and I.C. Eperon

Departments of Biochemistry and, University of Leicester University Road, Leicester LE1 7RH, UK *Department of Chemistry, University of Leicester University Road, Leicester LE1 7RH, UK

Received March 11, 1987. Revised April 29, 1987. Accepted April 29, 1987.

Transcription by T7 RNA polymerase has been studied using a chiral ATP analogue. The Sp diastereoisomer of adenosine 5'-0-(1-thiotriphos-phate) (ATP{alpha}S) was incorporated into RNA with an apparent KM of approximately 15 µM, similar to that for ATP; the Rp diastereoisomer was neither a substrate nor a competitive inhibitor. The configuration of the phosphodiester link in the RNA produced was analyzed with stereospecific nucleases. The rate of nuclease digestion was compared with the rate of digestion of phosphorothioate-substituted RNA of known stereochemistry synthesized by E.coli RNA polymerase. Surprisingly, the nucleases exhibited reduced discrimination compared with their activity on dinucleotides. The results show that phosphorothioate-substituted RNA transcribed by T7 RNA polymerase has the same configuration as that transcribed by E.coli RNA polymerase, ie. Rp. Thus, the reaction proceeds with inversion of configuration at phosphorus.


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
S. GOPALAKRISHNA, V. GUSTI, S. NAIR, S. SAHAR, and R. K. GAUR
Template-dependent incorporation of 8-N3AMP into RNA with bacteriophage T7 RNA polymerase
RNA, November 18, 2004; 10(11): 1820 - 1830.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
S. Krebs, I. Medugorac, D. Seichter, and M. Forster
RNaseCut: a MALDI mass spectrometry-based method for SNP discovery
Nucleic Acids Res., April 1, 2003; 31(7): e37 - e37.
[Abstract] [Full Text] [PDF]

Home page
 Eukaryot CellHome page
R. P. Igo Jr., D. S. Weston, N. L. Ernst, A. K. Panigrahi, R. Salavati, and K. Stuart
Role of Uridylate-Specific Exoribonuclease Activity in Trypanosoma brucei RNA Editing
Eukaryot. Cell, February 1, 2002; 1(1): 112 - 118.
[Abstract] [Full Text] [PDF]

Home page
 ScienceHome page
R. Padgett, M Podar, S. Boulanger, and P. Perlman
The stereochemical course of group II intron self-splicing
Science, December 9, 1994; 266(5191): 1685 - 1688.
[Abstract] [PDF]

Home page
 ScienceHome page
J. McSwiggen and T. Cech
Stereochemistry of RNA cleavage by the Tetrahymena ribozyme and evidence that the chemical step is not rate-limiting
Science, May 12, 1989; 244(4905): 679 - 683.
[Abstract] [PDF]

Home page
 ScienceHome page
J Rajagopal, J. Doudna, and J. Szostak
Stereochemical course of catalysis by the Tetrahymena ribozyme
Science, May 12, 1989; 244(4905): 692 - 694.
[Abstract] [PDF]


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.