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M9480468.TXT
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1994-08-20
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Document 0468
DOCN M9480468
TI Identification and analysis of the pseudoknot-containing gag-pro
ribosomal frameshift signal of simian retrovirus-1.
DT 9410
AU ten Dam E; Brierley I; Inglis S; Pleij C; Leiden Institute of Chemistry,
Department of Biochemistry,; Gorlaeus Laboratories, Leiden University,
The Netherlands.
SO Nucleic Acids Res. 1994 Jun 25;22(12):2304-10. Unique Identifier :
AIDSLINE MED/94310058
AB The pro and pol genes of simian retrovirus-1 (SRV-1) are expressed as
parts of a fusion protein generated by -1 ribosomal frameshifting. To
investigate the requirements for frameshifting at the gag-pro overlap,
we have inserted a stretch of 58 nucleotides containing the proposed
frameshift signal into a plasmid that allows monitoring of translation
in all three reading frames. In vitro translation of mRNAs derived from
this plasmid indicated that the 58 nucleotides from the SRV-1 gag-pro
overlap were sufficient to induce an efficient -1 shift in a
heterologous context. Mutational analysis demonstrated that the slip
site is formed at the heptanucleotide G GGA AAC. The frameshift
efficiency of the wild type sequence in rabbit reticulocyte lysate was
23%. A second component of the frameshift signal is formed by a
pseudoknot seven bases downstream of the slip site. The presence of this
pseudoknot was confirmed by mutational analysis, employing complementary
and compensatory base changes, and by probing the structure of short RNA
transcripts containing the frameshift signal. Adding increasing amounts
of an SRV-1 pseudoknot containing RNA transcript to a translation
reaction programmed with an SRV-1 frameshift reporter mRNA had no effect
on the frameshift efficiency, arguing against the role of a specific
pseudoknot-recognising factor in the frameshifting process.
DE Base Sequence Cloning, Molecular Frameshift Mutation *Gene Expression
Regulation, Viral *Genes, gag *Genes, pol Molecular Sequence Data
Nucleic Acid Conformation Regulatory Sequences, Nucleic Acid
Retroviruses Type D, Simian/*GENETICS RNA, Viral/CHEMISTRY/*GENETICS
Support, Non-U.S. Gov't Translation, Genetic Viral Proteins/GENETICS
JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).