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Recombinant RNase Z Does Not Recognize CCA as Part of the tRNA and Its Cleavage Efficieny Is Influenced by Acceptor Stem Length
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S. Schiffer
Published/Copyright:
June 1, 2005
Abstract
One of the essential maturation steps to yield functional tRNA molecules is the removal of 3'-trailer sequences by RNase Z. After RNase Z cleavage the tRNA nucleotidyl transferase adds the CCA sequence to the tRNA 3terminus, thereby generating the mature tRNA. Here we investigated whether a terminal CCA triplet as 3'-trailer or embedded in a longer 3'- trailer influences cleavage site selection by RNase Z using three activities: a recombinant plant RNase Z, a recombinant archaeal RNase Z and an RNase Z active wheat extract. A trailer of only the CCA trinucleotide is left intact by the wheat extract RNase Z but is removed by the recombinant plant and archaeal enzymes. Thus the CCA triplet is not recognized by the RNase Z enzyme itself, but rather requires cofactors still present in the extract. In addition, we investigated the influence of acceptor stem length on cleavage by RNase Z using variants of wildtype tRNATyr. While the wild type and the variant with 8 base pairs in the acceptor stem were processed efficiently by all three activities, variants with shorter and longer acceptor stems were poor substrates or were not cleaved at all.
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Published Online: 2005-06-01
Published in Print: 2003-03-14
Copyright © 2003 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
- Roumen Tsanev, Pioneer of the Early Days of Nucleic Acid Gel Electrophoresis and Histone Code Epigenetics
- Recombinant RNase Z Does Not Recognize CCA as Part of the tRNA and Its Cleavage Efficieny Is Influenced by Acceptor Stem Length
- HIV-1 TAR as Anchoring Site for Optimized Catalytic RNAs
- Transcription of Different Exons 1 of the Human Neuronal Nitric Oxide Synthase Gene Is Dynamically Regulated in a Cell- and Stimulus- Specific Manner
- Hepatitis B Virus Core Antigen: Enhancement of Its Production in Escherichia coli, and Interaction of the Core Particles with the Viral Surface Antigen
- Determinants of Enzymatic Specificity in the Cys-Met-Metabolism PLP-Dependent Enzyme Family: Crystal Structure of Cystathionine γ-Lyase from Yeast and Intrafamiliar Structure Comparison
- In vitro Uptake and Stability Study of pVEC and Its All-D Analog
- Calpastatin Exon 1B-Derived Peptide, a Selective Inhibitor of Calpain: Enhancing Cell Permeability by Conjugation with Penetratin
- Structure-Activity Relationships of Glucose-Dependent Insulinotropic Polypeptide (GIP)
- Interleukin-6 N-Terminal Peptides Modulate the Expression of junB Protooncogene and the Production of Fibrinogen in HepG2 Cells
- Induction of Apoptosis in Vascular Cells by Plasminogen Activator Inhibitor-1 and High Molecular Weight Kininogen Correlates with Their Anti-Adhesive Properties
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