Functional analysis of the SRV-1 RNA frameshifting pseudoknot
| Autor: | Cornelis W. Hilbers, Hans A. Heus, René C. L. Olsthoorn, Richard Reumerman, Cornelis W. A. Pleij |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Gene Expression Regulation
Viral Genetics Translational frameshift Base pair viruses Frameshifting Ribosomal RNA Regulatory Sequences Ribonucleic Acid Biology Slippery sequence Ribosome Regulatory sequence Mutation Biophysics Nucleic Acid Conformation RNA Viral Mason-Pfizer monkey virus Biophysical Chemistry Pseudoknot GeneralLiterature_REFERENCE(e.g. dictionaries encyclopedias glossaries) Triple helix |
| Zdroj: | Nucleic Acids Research, 38, 7665-7672 Nucleic Acids Research Nucleic Acids Research, 38, 21, pp. 7665-7672 |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkq629 |
| Popis: | Simian retrovirus type-1 uses programmed ribosomal frameshifting to control expression of the Gag-Pol polyprotein from overlapping gag and pol open-reading frames. The frameshifting signal consists of a heptanucleotide slippery sequence and a downstream-located 12-base pair pseudoknot. The solution structure of this pseudoknot, previously solved by NMR [Michiels,P.J., Versleijen,A.A., Verlaan,P.W., Pleij,C.W., Hilbers,C.W. and Heus,H.A. (2001) Solution structure of the pseudoknot of SRV-1 RNA, involved in ribosomal frameshifting. J. Mol. Biol., 310, 1109–1123] has a classical H-type fold and forms an extended triple helix by interactions between loop 2 and the minor groove of stem 1 involving base–base and base–sugar contacts. A mutational analysis was performed to test the functional importance of the triple helix for −1 frameshifting in vitro. Changing bases in L2 or base pairs in S1 involved in a base triple resulted in a 2- to 5-fold decrease in frameshifting efficiency. Alterations in the length of L2 had adverse effects on frameshifting. The in vitro effects were well reproduced in vivo, although the effect of enlarging L2 was more dramatic in vivo. The putative role of refolding kinetics of frameshifter pseudoknots is discussed. Overall, the data emphasize the role of the triple helix in −1 frameshifting. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|