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Mutational analysis of the RNA pseudoknot involved in efficient ribosomal frameshifting in simian retrovirus-1.
- Source :
-
Nucleic acids research [Nucleic Acids Res] 1998 Mar 15; Vol. 26 (6), pp. 1369-72. - Publication Year :
- 1998
-
Abstract
- Mutational effects on frameshifting efficiency of the RNA pseudoknot involved in ribosomal frameshifting in simian retrovirus-1 (SRV-1) have been investigated. The primary sequence and the proposed secondary structure of the SRV-1 pseudoknot are similar to those of other efficient frameshifting pseudoknots in mouse mammary tumor virus (MMTV) and feline immunodeficiency virus (FIV), where an unpaired adenine nucleotide intercalates between stem 1 and stem 2. In SRV-1 pseudoknot, the adenine nucleotide in between stem 1 and stem 2 has a potential to form an A*U base pair with the last uridine nucleotide in the loop 2, resulting in a continuous A-form helix with coaxially stacked stem 1 and stem 2. To test whether this A*U base pairing and coaxial stacking of stem 1 and stem 2 is absolutely required for efficient frameshifting in SRV-1, a series of mutants changing this potential A.U base pair to either G.C base pair or A.A, A.G, A.C, G.A, G.G mismatch is generated, and their frameshifting efficiencies are investigated in vitro using rabbit reticulocyte lysate translation assay. The frameshifting abilities of these mutant pseudoknots are similar to that of the wild-type pseudoknot, suggesting that the A*U base pair in between stem 1 and stem 2 is not necessary to promote efficient frameshifting in SRV-1. These results reveal that coaxial stacking of stem 1 and stem 2 with a Watson-Crick A.U base pair in between two stems is not a required structural feature of the pseudoknot for promoting efficient frameshifting in SRV-1. Our mutational data suggest that SRV-1 pseudoknot adopts similar structural features common to other efficient frameshifting pseudoknots as observed in MMTV and FIV.
- Subjects :
- Animals
Base Composition
Base Sequence
Cats
Immunodeficiency Virus, Feline genetics
In Vitro Techniques
Mammary Tumor Virus, Mouse genetics
Mice
Mutagenesis, Site-Directed
Nucleic Acid Conformation
Rabbits
Reticulocytes metabolism
Species Specificity
Frameshifting, Ribosomal
Mutation
RNA, Viral chemistry
RNA, Viral genetics
Retroviruses, Simian genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0305-1048
- Volume :
- 26
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Nucleic acids research
- Publication Type :
- Academic Journal
- Accession number :
- 9490779
- Full Text :
- https://doi.org/10.1093/nar/26.6.1369