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In vitro selection of RNA specifically c ... teriophage T4 RegB endonuclease
In vitro selection of RNA specifically cleaved by bacteriophage T4 RegB endonuclease.
54
T4 RegB endonuclease specifically cleaves at -GGAG- sites in several early T4 messages, rendering them nonfunctional. Not all -GGAG- sites are processed equally by RegB; those found at the Shine-Dalgarno sequences and in intercistronic regions are processed with higher efficiency than the -GGAG- sites located in coding regions. The low activity of RegB observed in vitro is enhanced by 1-2 orders of magnitude by the Escherichia coli ribosomal protein S1. We have used SELEX (systematic evolution of ligands by exponential enrichment) on a combinatorial RNA library to obtain molecules that are specifically cleaved by T4 RegB endonuclease in the presence of S1. The sequences obtained contain the required -GGAG- tetranucleotide and were unusually enriched in adenosine and cytosine nucleotides. No consensus structure or sequence motif other than -GGAG- was conserved among the selected molecules. The majority of the RNAs are entirely dependent on S1 for RegB-catalyzed cleavage; however, a few RNAs are found to be S1 independent but are cleaved by RegB with much lower rates.
Jayasena VK, Brown D, Shtatland T, Gold L
Biochemistry
1996-02-20 00:00
35
7
2349-56
Bacteriophage T4,Base Sequence,DNA Primers,Endoribonucleases,Hydrolysis,Molecular Sequence Data,Nucleic Acid Conformation,RNA,RNA Processing, Post-Transcriptional,Substrate Specificity,DNA Primers,RNA,Endoribonucleases
Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder 80309-0347, USA
Biochemistry
NIGMS GM 19963, NIGMS GM 28685
0006-2960
10.1021/bi951879b
bi951879b
411
True
8652576
