Effects of 3'-terminal phosphates in RNA produced by ribozyme cleavage

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Publication Type Journal Article
School or College School of Medicine
Department Human Genetics
Program Institute of Human Genetics; Howard Hughes Medical Institute (HHMI)
Creator Gesteland, Raymond F.; Alam, Steve L.; Nelson, Chad; Felden, Brice; Atkins, John F.
Title Effects of 3'-terminal phosphates in RNA produced by ribozyme cleavage
Date 1998
Description During the course of a structural investigation of a selenocysteine insertion element from rat Type 1 iodothyronine 59-deiodinase (D1 SECIS), we wished to prepare milligram quantities of an RNA that comprised the upper stem/loop of this RNA structure (41 nt; Fig+ 1A)+ This stem/loop is a stimulator required for the insertion of selenocysteins at a specific internal UGA stop codon (Berry et al+, 1991)+ Construction of the upper stem/ loop RNA required that the base of the stem end with four noncanonical base pairs (Walczak et al+, 1996), with no additional base pairing below this terminal quartet, as determined from in vitro assays (Martin et al+, 1998)+ This RNA has been produced synthetically for crystallization trials, so we sought a thorough comparison between this molecule and the RNA produced by run-off transcription+ Because the RNA to be studied begins with an adenosine and ends in a uridine, a ribozyme was used at each end to produce homogeneous RNA+ The system used to accomplish this has been described previously (Ferre-D™Amare & Doudna, 1996) and the resulting T7-produced RNA would differ from the synthetic RNA only by the 39- cyclic phosphate+ In making preliminary imino proton NMR assignments for both of these RNAs, we concluded that the base pairing for both RNAs is nearly identical, as evident from the one-dimensional NMR spectra collected in 90% H2O (Fig+ 2)+ Close inspection shows that there are subtle differences near the RNA helix end
Type Text
Publisher Cold Spring Harbor Lab Press
First Page 607
Last Page 612
Subject In vitro runoff transcription; RNA; Trans-acting ribozymes
Subject MESH Ribosomes; RNA; Proteins
Language eng
Bibliographic Citation Alam,S.L., Nelson, C.C., Felden, B., Atkins, J.F., and Gesteland, R.F. 1998. Effects of 3'-terminal phosphates in RNA produced by ribozyme cleavage. RNA 4:607-612
Rights Management (c) Cold Spring Harbor Laboratory Press-CSHL Press
Format Medium application/pdf
Format Extent 250,170 Bytes
Identifier ir-main,3738
ARK ark:/87278/s6988r40
Setname ir_uspace
Date Created 2012-06-13
Date Modified 2021-05-06
ID 702654
Reference URL https://collections.lib.utah.edu/ark:/87278/s6988r40