PMID:17623669
| Citation |
Waas, WF, Druzina, Z, Hanan, M and Schimmel, P (2007) Role of a tRNA base modification and its precursors in frameshifting in eukaryotes. J. Biol. Chem. 282:26026-34 |
|---|---|
| Abstract |
Little is known about the role of specific base modifications of transfer RNAs. Wyosine bases are tRNA(Phe)-specific modifications that are distinguished by differentiated, lateral side chains and base methylations appended to the core ring structure of a universally conserved G37, adjacent to the anticodon of Phe tRNAs. Based on previous data, we hypothesized that this modification was needed for -1 frameshifting. Using a reporter system incorporating a SCV-LA yeast virus slippery site for detecting -1 frameshifts in vivo, yeast strains were created that enabled chemical-genetic dissection of the role of different functional groups of wyebutosine that are added in a three-step post-transcriptional set of reactions. With this system, hypomodification increased Phe-specific frameshifting, with incremental changes in frameshift efficiency after specific intermediates in the progression of wyebutosine synthesis. These data combined with investigations of wild-type and hypomodified tRNA binding to ribosomes suggest that frameshift efficiency is kinetically and not thermodynamically controlled. The progressive nature of frameshift efficiency with the stage of modification is consistent with a stepwise evolution and tuning of frameshift potential. The stepwise tuning of frameshift efficiency could explain why tRNA(Phe) in some eukaryotes is not fully modified but, rather, hypomodified to capture a specific frameshift potential. |
| Links |
PubMed Online version:10.1074/jbc.M703391200 |
| Keywords |
Evolution, Molecular; Frameshifting, Ribosomal/genetics; Genes, Reporter/genetics; RNA Precursors/genetics; RNA Precursors/metabolism; RNA Processing, Post-Transcriptional/physiology; RNA, Fungal/genetics; RNA, Fungal/metabolism; RNA, Transfer, Phe/genetics; RNA, Transfer, Phe/metabolism; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/metabolism |
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